U.S. patent number 10,071,051 [Application Number 14/166,950] was granted by the patent office on 2018-09-11 for wt1 peptide cancer vaccine composition for transdermal administration.
This patent grant is currently assigned to NITTO DENKO CORPORATION, OSAKA UNIVERSITY. The grantee listed for this patent is NITTO DENKO CORPORATION, OSAKA UNIVERSITY. Invention is credited to Daisuke Asari, Mitsuhiko Hori, Yoshiki Maeda, Arimichi Okazaki, Katsuyuki Okubo, Takuya Shishido, Haruo Sugiyama.
United States Patent |
10,071,051 |
Okubo , et al. |
September 11, 2018 |
WT1 peptide cancer vaccine composition for transdermal
administration
Abstract
The present invention provides a cancer vaccine composition for
transdermal administration for cellular immunity induction,
comprising (i) a WT1 peptide and/or a modified WT1 peptide; and
(ii) a pharmacologically acceptable acid as a first cellular
immunity induction promoter, or a pharmacologically acceptable salt
thereof.
Inventors: |
Okubo; Katsuyuki (Osaka,
JP), Maeda; Yoshiki (Osaka, JP), Okazaki;
Arimichi (Osaka, JP), Asari; Daisuke (Osaka,
JP), Shishido; Takuya (Osaka, JP), Hori;
Mitsuhiko (Osaka, JP), Sugiyama; Haruo (Osaka,
JP) |
Applicant: |
Name |
City |
State |
Country |
Type |
NITTO DENKO CORPORATION
OSAKA UNIVERSITY |
Osaka
Osaka |
N/A
N/A |
JP
JP |
|
|
Assignee: |
NITTO DENKO CORPORATION (Osaka,
JP)
OSAKA UNIVERSITY (Osaka, JP)
|
Family
ID: |
50028730 |
Appl.
No.: |
14/166,950 |
Filed: |
January 29, 2014 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20140220057 A1 |
Aug 7, 2014 |
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Foreign Application Priority Data
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Feb 5, 2013 [JP] |
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2013-020908 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/045 (20130101); A61K 9/0014 (20130101); A61K
31/716 (20130101); A61K 39/0011 (20130101); A61K
39/001153 (20180801); A61K 31/739 (20130101); A61P
35/00 (20180101); A61K 31/437 (20130101); A61K
38/10 (20130101); A61P 37/04 (20180101); A61K
31/708 (20130101); A61K 31/513 (20130101); A61K
31/728 (20130101); A61K 31/451 (20130101); A61K
2039/54 (20130101) |
Current International
Class: |
A61K
9/00 (20060101); A61K 31/728 (20060101); A61K
31/739 (20060101); A61K 38/10 (20060101); A61K
39/00 (20060101); A61K 31/045 (20060101); A61K
31/437 (20060101); A61K 31/451 (20060101); A61K
31/513 (20060101); A61K 31/708 (20060101); A61K
31/716 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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2228072 |
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Sep 2010 |
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EP |
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7-505883 |
|
Jun 1995 |
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JP |
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2002-531415 |
|
Sep 2002 |
|
JP |
|
2007-529531 |
|
Oct 2007 |
|
JP |
|
200874763 |
|
Apr 2008 |
|
JP |
|
2008127277 |
|
Jun 2008 |
|
JP |
|
4422903 |
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Dec 2009 |
|
JP |
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2014169280 |
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Sep 2014 |
|
JP |
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2192884 |
|
Nov 2002 |
|
RU |
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93/20847 |
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Oct 1993 |
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WO |
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00/32228 |
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Jun 2000 |
|
WO |
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2003106682 |
|
Dec 2003 |
|
WO |
|
2005/087238 |
|
Sep 2005 |
|
WO |
|
2008093772 |
|
Aug 2008 |
|
WO |
|
Other References
Inoue et al. (Journal of Investigative Dermatology, 2007,
127:614-621). cited by examiner .
Karande et al. (Annual Rev. Chem. Biomol. Eng., 2010, 1:175-201).
cited by examiner .
Lim et al. Int. Immunopharmacol. Jan. 2003; 3(1): 115-118. cited by
examiner .
Extended European search report issued with respect to European
application No. 14000317.9, dated Apr. 7, 2014. cited by applicant
.
Yoshihiro Oka et al., Current Opinion in Immunology, 20: 211-220
(2008). cited by applicant .
Hosoi Akihiro et al., Cancer Research, 68, 2008, pp. 3941-3949.
cited by applicant .
Zhengrong Cui et al., Pharmaceutical Research, vol. 19, No. 7,
2002, pp. 947-953. cited by applicant .
European Office Action issued with respect to European Application
No. 14000317.9, dated Dec. 15, 2016. cited by applicant .
Chinese Office Action issued in Patent Application No.
201410042896.X, dated Mar. 1, 2017, along with an English
translation thereof. cited by applicant .
European Search Report from Application No. 14000317.9 dated Jan.
12, 2018. cited by applicant .
Japanese Office Action from Application No. 2014-014800 dated Oct.
31, 2017. cited by applicant .
Russian Office Action and Search Report from Application No.
2014102940 dated Dec. 26, 2017. cited by applicant .
Chinese Office Action from Application No. 201410042896.X dated
Jan. 19, 2018. cited by applicant .
Karande, Pankaj, "Transcutaneous immunization using common
chemicals", 2009, Journal of Controlled Release, pp. 134-140, vol.
138. cited by applicant.
|
Primary Examiner: Gamett; Daniel C
Attorney, Agent or Firm: Greenblum & Bernstein,
P.L.C.
Claims
What is claimed is:
1. A method for inducing cellular immunity in a subject, which
comprises transdermally administering to the subject a cancer
vaccine composition comprising: (i) a WT1 peptide and/or a modified
WT1 peptide; and (ii) a first cellular immunity induction promoter
that is a pharmacologically acceptable acid or a pharmacologically
acceptable salt thereof, wherein the pharmacologically acceptable
acid is selected from the group consisting of isostearic acid,
decanoic acid, myristic acid, and malic acid.
2. The method according to claim 1, wherein the method is for
treating a cancer in the subject.
3. The method according to claim 1, wherein the cancer vaccine
composition further comprises a second cellular immunity induction
promoter selected from a TLR ligand, a cyclic dinucleotide, a
helper peptide, an immunomodulatory small molecule drug, a
cyclooxygenase inhibitor, a prostaglandin receptor antagonist, a
prostaglandin receptor agonist, a TSLP production inhibitor, an
adenylate cyclase inhibitor, an omega-3 fatty acid, a PPAR agonist,
a dopamine receptor antagonist, a dopamine receptor agonist, a
histamine receptor agonist, a histamine receptor antagonist, a
serotonin receptor agonist, a serotonin receptor antagonist, a
vasopressin receptor antagonist, a vasopressin receptor agonist, a
muscarine receptor antagonist, a muscarine receptor agonist, an
adrenalin receptor antagonist, an adrenalin receptor agonist, an
angiotensin receptor agonist, a GABA receptor agonist, a thrombin
receptor antagonist, a thrombin receptor agonist, an opioid
receptor agonist, an ADP receptor agonist, a leukotriene receptor
antagonist, a leukotriene receptor agonist, a melatonin receptor
agonist, a somatostatin receptor agonist, a cannabinoid receptor
agonist, a sphingosine-1 phosphate receptor agonist, a metabotropic
glutamate receptor agonist, a phospholipase A2 inhibitor, a
TGF-.beta. production inhibitor, and a Th2 cytokine inhibitor, and
a combination of two or more kinds of them.
4. The method according to claim 3, wherein the second cellular
immunity induction promoter is a helper peptide.
5. The method according to claim 3, wherein the second cellular
immunity induction promoter is a combination of a helper peptide
and at least one substance selected from the group consisting of a
TLR ligand, a cyclic dinucleotide, an immunomodulatory small
molecule drug, a cyclooxygenase inhibitor, a prostaglandin receptor
antagonist, a prostaglandin receptor agonist, a TSLP production
inhibitor, an adenylate cyclase inhibitor, an omega-3 fatty acid, a
PPAR agonist, a dopamine receptor antagonist, a dopamine receptor
agonist, a histamine receptor agonist, a histamine receptor
antagonist, a serotonin receptor agonist, a serotonin receptor
antagonist, a vasopressin receptor antagonist, a vasopressin
receptor agonist, a muscarine receptor antagonist, a muscarine
receptor agonist, an adrenalin receptor antagonist, an adrenalin
receptor agonist, an angiotensin receptor agonist, a GABA receptor
agonist, a thrombin receptor antagonist, a thrombin receptor
agonist, an opioid receptor agonist, an ADP receptor agonist, a
leukotriene receptor antagonist, a leukotriene receptor agonist, a
melatonin receptor agonist, a somatostatin receptor agonist, a
cannabinoid receptor agonist, a sphingosine-1 phosphate receptor
agonist, a metabotropic glutamate receptor agonist, a phospholipase
A2 inhibitor, a TGF-.beta. production inhibitor, and a Th2 cytokine
inhibitor.
6. The method according to claim 1, wherein the cancer vaccine
composition is in the form of a cream formulation.
7. The method according to claim 1, wherein the cancer vaccine
composition is in the form of a liquid formulation.
8. The method according to claim 1, wherein the cancer vaccine
composition is administered under a mildly irritating
condition.
9. The method according to claim 8, wherein the mildly irritating
condition is a condition under which transepidermal water loss
(TEWL) in a model animal for skin irritation evaluation before the
administration of the composition is 50 g/hm or less.
10. The method according to claim 8, wherein the mildly irritating
condition is a condition under which the cutaneous TSLP level in a
model animal for skin irritation evaluation at completion of the
administration of the composition is 10000 pg/mg protein or
less.
11. The method according to claim 1, wherein the pharmacologically
acceptable acid or a pharmacologically acceptable salt thereof is
isostearic acid.
Description
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been
submitted electronically in ASCII format and is hereby incorporated
by reference in its entirety. Said ASCII copy, created on Mar. 18,
2014, is named P45215_SL.txt and is 3,362 bytes in size.
TECHNICAL FIELD
The present invention relates to a cancer vaccine for transdermal
administration comprising a WT1 peptide and/or a modified WT1
peptide, and a cellular immunity induction promoter.
BACKGROUND ART
There are a cancer vaccine that prevents virus infection to prevent
a cancer caused by the virus, and a cancer vaccine which provides
the result that cancer cells are specifically attacked by the
immune system via the recognition of a cancer-specific antigen by
the immune mechanism, particularly, the cellular immune mechanism
in which cytotoxic T cells (CTL) play an important role. The former
is not effective at all for a cancer in which the virus does not
participate. The latter is a cancer therapeutic strategy of
targeting an antigen possessed by a cancer cell itself. It is
considered that the latter is widely effective for cancers having
antigen by specifying the antigen. Inter alia, a cancer vaccine
based on the viewpoint of the latter can treat tumors that are
difficult to remove by surgical operation because of their size,
and causes less side effects as compared with the conventional
therapies such as chemotherapy and radiation therapy.
WT1 (Wilm's tumor 1) gene is overexpressed in many hematopoietic
tumors and solid cancers, for example, acute myelocytic leukemia,
acute lymphocytic leukemia, chronic myelocytic leukemia,
myelodysplastic syndrome, multiple myeloma, non-Hodgkin's lymphoma,
lung cancer, breast cancer, stomach cancer, large intestine/rectum
cancer, pancreas cancer, bile duct cancer, head and neck squamous
cell cancer, thyroid cancer, kidney cancer, prostate cancer,
ovarian cancer, uterine cancer, bone soft tissue sarcoma, malignant
melanoma, malignant mesothelioma, testicular germ cell tumor and
malignant glioma. Those cancers overproduce the WT1 protein. The
WT1 protein is fragmented in the cancer cell to produce partial
peptides consisting of 8 to 12 amino acids. A WT1 peptide is one of
the peptide fragment which has been bound with the MHC class I
molecule in a cancer cell, moved to the surface of the cancer cell,
and presented as an antigen bound to the MHC class I molecule on
the cancer cell surface. The WT1 peptide becomes a mark of the
cancer cell. The amino acid sequence of the WT1 peptide conforms to
the type of the MHC class I molecule of the cell. For example, in
the case of a cell having HLA-A*0201-type MHC, a HLA-A*0201-type
MHC restricted WT1 peptide such as Db126 peptide consisting of 9
amino acids is generated, and in the case of a cell having
HLA-A*2402-type MHCa HLA-A*2402-type MHC restricted WT1 peptide
such as Db235 peptide consisting of 9 amino acids is generated. In
the case of a cell having other MHC, such as HLA-A26 type (WO
2005/095598), HLA-A*3303 type (WO 2007/097358), or HLA-A*1101 type
(WO 2008/081701), each MHC restricted WT1 peptide is generated.
When a WT1 peptide, or a modified WT1 peptide in which a part of
amino acids of the WT1 peptide is substituted or modified is
administered to a living body as an antigen (herein, a WT1 peptide
or a modified WT1 peptide which has been administered as an antigen
is referred to as "WT1 antigen peptide"), the WT1 antigen peptide
is bound to the MHC Class I molecule on the surface of a dendritic
cell which is an antigen presenting cell, or the WT1 antigen
peptide is once taken into a dendritic cell, bound to the MHC class
I molecule of the dendritic cell and then, is moved to the surface
of the dendritic cell, thereby, is presented as an antigen bound to
the MHC class I molecule on the surface of the dendritic cell. An
activated dendritic cell having the WT1 antigen peptide/MHC class I
molecule complex is moved to the regional lymph node, and activates
a CD8-positive T lymphocyte which recognizes the WT1 antigen
peptide/MHC class I molecule complex to differentiate and
proliferate the cell into a cytotoxic T cell (CTL). CTL recognizes
tumor cells having the complex of a WT1 peptide (derived from the
endogenous WT1 protein) of the same amino acid sequence as the WT1
antigen peptide and the MHC class I molecule, or a tumor cell
having a complex of a WT1 peptide (derived from the endogenous WT1
protein) of an amino acid sequence having cross immunoreactivity
with the WT1 antigen peptide and the MHC class I molecule, and
attacks the recognized tumor cells. Therefore, the aforementioned
various MHC restricted WT1 peptides such as Db126 peptide and Db235
peptide, and modified WT1 peptides in which a part of amino acids
of them are substituted or modified are useful as cancer vaccines
(Non-Patent Document 1).
It is also known that an adjuvant is utilized in order to enhance
the action as cancer vaccine of the WT1 peptide and/or the modified
WT1 peptide. As the adjuvant for the WT1 peptide and/or the
modified WT1 peptide, for example, mineral gels such as aluminum
hydroxide; surfactants such as lysolecithin, and pluronicpolyol;
polyanions; peptides; or oil emulsions (Patent Document 1), and
GM-CSF, BCG-CWS and Montanide ISA51 (Non-Patent Document 1) are
known. In addition to them, a variety of vaccine adjuvants
including cyclic dinucleotide analogs (Patent Document 3 and Patent
Document 4) such as 1H-imidazo[4,5-c]quinoline-4-amine, imiquimod
(Patent Document 2), and cyclic di-GMP (c-di-GMP), and TLR2, 3, 7,
8 and 9 ligands (Patent Document 5) have been known. In addition,
it is also known that immunity induced by transdermal
administration of imiquimod-containing peptide is further enhanced
by adding Peptide-25 (Non-Patent Document 2).
In general, vaccines are administered by subcutaneous or
intradermal injection. In addition to those routes, immunity
induction by a variety of administration routes, for example,
transdermal administration (Patent Document 5 and Non-Patent
Document 2), buccal administration, nasal administration, and
sublingual administration (Non-Patent Document 3, Patent Document
6, and Patent Document 7) have been tried.
LIST OF DOCUMENTS
[Patent Document 1] Japanese Patent No. 4422903 [Patent Document 2]
JP 7-505883 A [Patent Document 3] JP 2007-529531 A [Patent Document
4] US Patent Application Publication No. 2008/0286296 [Patent
Document 5] US Patent Application Publication No. 2008/0193487
[Patent Document 6] JP 2002-531415 A [Patent Document 7] US Patent
Application Publication No. 2008/0112974
Non-Patent Documents
[Non-Patent Document 1] Yoshihiro Oka et al., Current Opinion in
Immunology, 20: 211-220 (2008) [Non-Patent Document 2] Hosoi
Akihiro et al., Cancer Research, 68, 3941-3949 (2008) [Non-Patent
Document 3] Zhengrong Cui et al., Pharmaceutical Research, Vol. 19,
No. 7, 947-953 (2002)
SUMMARY OF THE INVENTION
It is well-known that an adjuvant is used to enhance efficacy of a
vaccine. Suitable adjuvants generally vary depending on, for
example, the kind of the antigen, the administration route, and the
immune response which is desired to be induced (i.e. cellular
immunity or humoral immunity). Further, in addition to the
adjuvant, there are a variety of substances which promote the
induction of the immunity. Then, an object of the present invention
is to provide a composition for use as a cancer vaccine with higher
efficacy and is convenient for use.
A microorganism or a virus itself, or a part of them is contained
in a widely used vaccine and the vaccine is administered to induce
immune response. Usually, since invasion of the microorganism or
virus is inhibited by the skin due to the size thereof, it is
necessary that the vaccine is invasively administered into the
body. Therefore, vaccines are usually administered by injection.
However, the injection has some problems including pain, fear,
injection scar, and subsequent scarring cicatrization. People other
than health care workers are not permitted to perform the
injection. Intradermal injection which can introduce higher immune
response is a difficult administration technique. There is a risk
of accidental infection of the health care workers due to
needlestick injury. Patients are needed to visit the hospital
repeatedly when administration is performed repetitively. Medical
wastes which necessitate special disposition such as injection
needles are generated. In view of the above issues, injection is
not necessarily the optimal administration route.
A WT1 peptide and/or a modified WT1 peptide can activate CTL
(cytotoxic T cell) via a MHC class I molecule, that is, the peptide
can induce cellular immunity. The WT1 peptide and/or the modified
WT1 peptide are a molecule having a molecular weight of about 700
to about 1600 and consisting of 8 to 12 amino acids, and are
significantly smaller than microorganisms or virus itself although
they are not considered as a small-molecule substance. It may be
possible that they are administered by a route other than
injection. However, a preparation for the administration of the
peptide vaccine in a rout other than injection has not been
developed yet. The reason includes many things, for example: a
suitable substance that can promote to induce the cellular immunity
has been unknown; it has also been unknown whether or not an
antigen can be delivered to a tissue suitable for the induction of
the cellular immunity. Inter alia, a substance that can promote to
induce the cellular immunity, which can be used with the antigen
when administered in a route other than injection has been
unknown.
It was found out that, by transdermal administration, the cellular
immunity induction effect which is equivalent to, or exceeds that
of injection can be obtained. Then, a substance suitable for
cellular immunity induction by transdermal administration of the
WT1 peptide and/or the modified WT1 peptide was searched and, as a
result, it was found out that use of a pharmacologically acceptable
acid or a pharmacologically acceptable salt thereof is suitable.
Further, it was found out that joint use of a pharmacologically
acceptable acid with one or more kinds of cellular immunity
induction promoters selected from the group consisting of a TLR
ligand, a cyclic dinucleotide, a helper peptide, an
immunomodulatory small molecule drug, a cyclooxygenase inhibitor, a
prostaglandin receptor antagonist, a prostaglandin receptor
agonist, a TSLP production inhibitor, an adenylate cyclase
inhibitor, an omega-3 fatty acid, a PPAR agonist, a dopamine
receptor antagonist, a dopamine receptor agonist, a histamine
receptor agonist, a histamine receptor antagonist, a serotonin
receptor agonist, a serotonin receptor antagonist, a vasopressin
receptor antagonist, a vasopressin receptor agonist, a muscarine
receptor antagonist, a muscarine receptor agonist, an adrenalin
receptor antagonist, an adrenalin receptor agonist, an angiotensin
receptor agonist, a GABA receptor agonist, a thrombin receptor
antagonist, a thrombin receptor agonist, an opioid receptor
agonist, an ADP receptor agonist, a leukotriene receptor
antagonist, a leukotriene receptor agonist, a melatonin receptor
agonist, a somatostatin receptor agonist, a cannabinoid receptor
agonist, a sphingosine-1 phosphate receptor agonist, a metabotropic
glutamate receptor agonist, a phospholipase A2 inhibitor, a
TGF-.beta. production inhibitor, and a Th2 cytokine inhibitor is
suitable. In a particularly suitable aspect, cellular immunity was
remarkably enhanced by a combination of a pharmacologically
acceptable acid and a TLR ligand, or a combination of a
pharmacologically acceptable acid, a TLR ligand and a helper
peptide. Further, it was found out that, by administration under
the mildly irritating condition, the high cellular immunity
inducing effect is obtained. Specifically, the high cellular
immunity inducing effect is obtained by selecting the mildly
irritating state where the transepidermal water loss (TEWL)
(g/hm.sup.2) which is an index of a skin of a model animal for skin
irritation evaluation before administration of a cancer vaccine
composition for transdermal administration is 50 or less, and
administering a cancer vaccine composition for transdermal
administration. Alternatively, the high cellular immunity inducing
effect is also obtained by rendering skin irritation property of a
cancer vaccine composition for transdermal administration such a
mildly irritating property that the cutaneous TSLP level (pg/mg
protein) of a model animal for skin irritation evaluation at
completion of administration becomes 10000 or less.
Therefore, the present invention, in a first aspect, provides
aspects listed below:
(1) A cancer vaccine composition for transdermal administration for
cellular immunity induction, comprising:
(i) a WT1 peptide and/or a modified WT1 peptide; and
(ii) a pharmacologically acceptable acid as a first cellular
immunity induction promoter or a pharmacologically acceptable salt
thereof;
(2) The cancer vaccine composition for transdermal administration
according to (1), wherein the pharmacologically acceptable acid or
a pharmacologically acceptable salt thereof is an organic acid or a
pharmacologically acceptable salt thereof;
(3) The cancer vaccine composition for transdermal administration
according to (2), wherein the organic acid or a pharmacologically
acceptable salt thereof is an organic compound containing carboxyl
group or an organic compound containing sulfonate group, or a
pharmacologically acceptable salt thereof; (4) The cancer vaccine
composition for transdermal administration according to (2),
wherein the organic acid or a pharmacologically acceptable salt
thereof is saturated or unsaturated straight or branched fatty acid
in which a saturated straight chain part has 8 to 20 carbon atoms,
lactic acid, malic acid, salicylic acid, maleic acid, citric acid,
or an organic compound containing sulfonate group, or a
pharmacologically acceptable salt thereof; (5) The cancer vaccine
composition for transdermal administration according to (2),
wherein the organic acid or a pharmacologically acceptable salt
thereof is fatty acid selected from the group consisting of
decanoic acid, lauric acid, myristic acid, isostearic acid and
oleic acid, or lactic acid, salicylic acid, citric acid or
methanesulfonic acid, or a pharmacologically acceptable salt
thereof; (6) The cancer vaccine composition for transdermal
administration according to any one of (1) to (5), further
comprising a second cellular immunity induction promoter selected
from a TLR ligand, a cyclic dinucleotide, a helper peptide, an
immunomodulatory small molecule drug, a cyclooxygenase inhibitor, a
prostaglandin receptor antagonist, a prostaglandin receptor
agonist, a TSLP production inhibitor, an adenylate cyclase
inhibitor, an omega-3 fatty acid, a PPAR agonist, a dopamine
receptor antagonist, a dopamine receptor agonist, a histamine
receptor agonist, a histamine receptor antagonist, a serotonin
receptor agonist, a serotonin receptor antagonist, a vasopressin
receptor antagonist, a vasopressin receptor agonist, a muscarine
receptor antagonist, a muscarine receptor agonist, an adrenalin
receptor antagonist, an adrenalin receptor agonist, an angiotensin
receptor agonist, a GABA receptor agonist, a thrombin receptor
antagonist, a thrombin receptor agonist, an opioid receptor
agonist, an ADP receptor agonist, a leukotriene receptor
antagonist, a leukotriene receptor agonist, a melatonin receptor
agonist, a somatostatin receptor agonist, a cannabinoid receptor
agonist, a sphingosine-1 phosphate receptor agonist, a metabotropic
glutamate receptor agonist, a phospholipase A2 inhibitor, a
TGF-.beta. production inhibitor, and a Th2 cytokine inhibitor, and
a combination of two or more kinds of them; (7) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a TLR
ligand; (8) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is a cyclic dinucleotide; (9) The
cancer vaccine composition for transdermal administration according
to (6), wherein the second cellular immunity induction promoter is
an immunomodulatory small molecule drug; (10) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
cyclooxygenase inhibitor; (11) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a prostaglandin receptor
antagonist and, further, the prostaglandin receptor antagonist is
an EP2 receptor antagonist, an EP4 receptor antagonist, a DP
receptor antagonist, or an IP receptor antagonist; (12) The cancer
vaccine composition for transdermal administration according to
(6), wherein the second cellular immunity induction promoter is a
prostaglandin receptor agonist and, further, the prostaglandin
receptor agonist is an EP3 receptor agonist; (13) The cancer
vaccine composition for transdermal administration according to
(6), wherein the second cellular immunity induction promoter is a
TSLP production inhibitor; (14) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is an adenylate cyclase
inhibitor; (15) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is an omega-3 fatty acid; (16) The
cancer vaccine composition for transdermal administration according
to (6), wherein the second cellular immunity induction promoter is
a PPAR agonist and, further, the PPAR agonist is a PPAR-.alpha.
agonist, a PPAR-.delta. agonist, or a PPAR-.gamma. agonist; (17)
The cancer vaccine composition for transdermal administration
according to (6), wherein the second cellular immunity induction
promoter is a dopamine receptor antagonist and, further, the
dopamine receptor antagonist is a D1 receptor antagonist, or a D5
receptor antagonist; (18) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a dopamine receptor agonist
and, further, the dopamine receptor agonist is a D2 receptor
agonist, a D3 receptor agonist, or a D4 receptor agonist; (19) The
cancer vaccine composition for transdermal administration according
to (6), wherein the second cellular immunity induction promoter is
a histamine receptor antagonist and, further, the histamine
receptor antagonist is a H1 receptor antagonist, or a H2 receptor
antagonist; (20) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is a histamine receptor agonist and,
further, the histamine receptor agonist is a H1 receptor agonist, a
H3 receptor agonist, or a H4 receptor agonist; (21) The cancer
vaccine composition for transdermal administration according to
(6), wherein the second cellular immunity induction promoter is a
serotonin receptor antagonist and, further, the serotonin receptor
antagonist is a 5-HT2 receptor antagonist, a 5-HT4 receptor
antagonist, a 5-HT6 receptor antagonist, or a 5-HT7 receptor
antagonist; (22) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is a serotonin receptor agonist and,
further, the serotonin receptor agonist is a 5-HT1 receptor
agonist, or a 5-HT2 receptor agonist; (23) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
vasopressin receptor antagonist and, further the vasopressin
receptor antagonist is a V2 receptor antagonist; (24) The cancer
vaccine composition for transdermal administration according to
(6), wherein the second cellular immunity induction promoter is a
vasopressin receptor agonist and, further, the vasopressin receptor
agonist is a V1 receptor agonist; (25) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
muscarine receptor antagonist and, further, the muscarine receptor
antagonist is a M1 receptor antagonist, a M3 receptor antagonist,
or a M5 receptor antagonist; (26) The cancer vaccine composition
for transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a muscarine receptor
agonist and, further, the muscarine receptor agonist is a M1
receptor agonist, a M2 receptor agonist, a M3 receptor agonist, a
M4 receptor agonist, or a M5 receptor agonist; (27) The cancer
vaccine composition for transdermal administration according to
(6), wherein the second cellular immunity induction promoter is an
adrenalin receptor antagonist and, further, the adrenalin receptor
antagonist is an .alpha.1 receptor antagonist, a .beta.1 receptor
antagonist, a .beta.2 receptor antagonist or a .beta.3 receptor
antagonist; (28) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is an adrenalin receptor agonist and,
further, the adrenalin receptor agonist is an .alpha.1 receptor
agonist, or an .alpha.2 receptor agonist; (29) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is an
angiotensin receptor agonist and, further, the angiotensin receptor
agonist is an AT2 receptor agonist; (30) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a GABA
receptor agonist and, further, the GABA receptor agonist is a
GABA.sub.B receptor agonist; (31) The cancer vaccine composition
for transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a thrombin receptor
antagonist and, further the thrombin receptor antagonist is a PAR-1
receptor antagonist; (32) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a thrombin receptor agonist
and, further the thrombin receptor agonist is a PAR-1 receptor
agonist; (33) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is an opioid receptor agonist; (34) The
cancer vaccine composition for transdermal administration according
to (6), wherein the second cellular immunity induction promoter is
a leukotriene receptor antagonist and, further, the leukotriene
receptor antagonist is a CysLT1 receptor antagonist, or a CysLT2
receptor antagonist; (35) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a leukotriene receptor
agonist and, further, the leukotriene receptor agonist is a BLT
receptor agonist; (36) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a melatonin receptor
agonist; (37) The cancer vaccine composition for transdermal
administration according to (6), wherein the second cellular
immunity induction promoter is a somatostatin receptor agonist;
(38) The cancer vaccine composition for transdermal administration
according to (6), wherein the second cellular immunity induction
promoter is a cannabinoid receptor agonist; (39) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
sphingosine-1 phosphate receptor agonist; (40) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
metabotropic glutamate receptor agonist and, further, the
metabotropic glutamate receptor agonist is an mGluR2 receptor
agonist, an mGluR3 receptor agonist, an mGluR4 receptor agonist, an
mGluR6 receptor agonist, an mGluR7 receptor agonist, or an mGluR8
receptor agonist; (41) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is an ADP receptor agonist;
(42) The cancer vaccine composition for transdermal administration
according to (6), wherein the second cellular immunity induction
promoter is a phospholipase A2 inhibitor; (43) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a
TGF-.beta. production inhibitor; (44) The cancer vaccine
composition for transdermal administration according to (6),
wherein the second cellular immunity induction promoter is a Th2
cytokine inhibitor; (45) The cancer vaccine composition for
transdermal administration according to (6), wherein the second
cellular immunity induction promoter is a helper peptide; (46) The
cancer vaccine composition for transdermal administration according
to (6), wherein the second cellular immunity induction promoter is
a combination of one or more kinds selected from the group
consisting of a TLR ligand, a cyclic dinucleotide, an
immunomodulatory small molecule drug, a cyclooxygenase inhibitor, a
prostaglandin receptor antagonist, a prostaglandin receptor
agonist, a TSLP production inhibitor, an adenylate cyclase
inhibitor, an omega-3 fatty acid, a PPAR agonist, a dopamine
receptor antagonist, a dopamine receptor agonist, a histamine
receptor agonist, a histamine receptor antagonist, a serotonin
receptor agonist, a serotonin receptor antagonist, a vasopressin
receptor antagonist, a vasopressin receptor agonist, a muscarine
receptor antagonist, a muscarine receptor agonist, an adrenalin
receptor antagonist, an adrenalin receptor agonist, an angiotensin
receptor agonist, a GABA receptor agonist, a thrombin receptor
antagonist, a thrombin receptor agonist, an opioid receptor
agonist, an ADP receptor agonist, a leukotriene receptor
antagonist, a leukotriene receptor agonist, a melatonin receptor
agonist, a somatostatin receptor agonist, a cannabinoid receptor
agonist, a sphingosine-1 phosphate receptor agonist, a metabotropic
glutamate receptor agonist, a phospholipase A2 inhibitor, a
TGF-.beta. production inhibitor, and a Th2 cytokine inhibitor, with
a helper peptide; (47) The cancer vaccine composition for
transdermal administration according to any one of (1) to (46),
which is in a form of a cream formulation; (48) The cancer vaccine
composition for transdermal administration according to any one of
(1) to (46), which is in a form of a liquid formulation; (49) The
cancer vaccine composition for transdermal administration according
to any one of (1) to (48), which is administered under a mildly
irritating condition; (50) The cancer vaccine composition for
transdermal administration according to (49), wherein the mildly
irritating condition is a condition under which transepidermal
water loss (TEWL) before administration in a model animal for skin
irritation evaluation is 50 g/hm.sup.2 or less; and (51) The cancer
vaccine composition for transdermal administration according to
(49) or (50), wherein the mildly irritating condition is a
condition under which the cutaneous TSLP level at completion of
administration in a model animal for skin irritation evaluation is
10000 pg/mg protein or less.
In other aspect, the cancer vaccine of the present invention can be
used for treating or preventing a cancer. Therefore, the present
invention also provides aspects listed below:
(52) A method of treating or preventing a cancer comprising
transdermally administering a therapeutically effective amount of
(i) a WT1 peptide and/or a modified WT1 peptide, and (ii) a
pharmacologically acceptable acid or a pharmacologically acceptable
salt thereof to a subject; (53) A method of treating or preventing
a cancer comprising transdermally administering, to a subject, a
therapeutically effective amount of (i) a WT1 peptide and/or a
modified WT1 peptide, (ii) a pharmacologically acceptable acid as a
first cellular immunity induction promoter, or a pharmacologically
acceptable salt thereof, and (iii) a second cellular immunity
induction promoter selected from a TLR ligand, a cyclic
dinucleotide, a helper peptide, an immunomodulatory small molecule
drug, a cyclooxygenase inhibitor, a prostaglandin receptor
antagonist, a prostaglandin receptor agonist, a TSLP production
inhibitor, an adenylate cyclase inhibitor, an omega-3 fatty acid, a
PPAR agonist, a dopamine receptor antagonist, a dopamine receptor
agonist, a histamine receptor agonist, a histamine receptor
antagonist, a serotonin receptor agonist, a serotonin receptor
antagonist, a vasopressin receptor antagonist, a vasopressin
receptor agonist, a muscarine receptor antagonist, a muscarine
receptor agonist, an adrenalin receptor antagonist, an adrenalin
receptor agonist, an angiotensin receptor agonist, a GABA receptor
agonist, a thrombin receptor antagonist, a thrombin receptor
agonist, an opioid receptor agonist, an ADP receptor agonist, a
leukotriene receptor antagonist, a leukotriene receptor agonist, a
melatonin receptor agonist, a somatostatin receptor agonist, a
cannabinoid receptor agonist, a sphingosine-1 phosphate receptor
agonist, a metabotropic glutamate receptor agonist, a phospholipase
A2 inhibitor, a TGF-.beta. production inhibitor, a Th2 cytokine
inhibitor, and a combination of two or more kinds of them; (54) A
method of treating or preventing a cancer comprising administering
a therapeutically effective amount of the cancer vaccine
composition for transdermal administration according to any one of
(1) to (51) to a subject; (55) A pharmacologically acceptable acid
or a pharmacologically acceptable salt thereof, for use as a
cellular immunity induction promoter for transdermally
administering a WT1 peptide and/or a modified WT1 peptide; and (56)
A combination of (i) a pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof, and (ii) one or more
kinds of substances selected from the group consisting of a TLR
ligand, a cyclic dinucleotide, a helper peptide, an
immunomodulatory small molecule drug, a cyclooxygenase inhibitor, a
prostaglandin receptor antagonist, a prostaglandin receptor
agonist, a TSLP production inhibitor, an adenylate cyclase
inhibitor, an omega-3 fatty acid, a PPAR agonist, a dopamine
receptor antagonist, a dopamine receptor agonist, a histamine
receptor agonist, a histamine receptor antagonist, a serotonin
receptor agonist, a serotonin receptor antagonist, a vasopressin
receptor antagonist, a vasopressin receptor agonist, a muscarine
receptor antagonist, a muscarine receptor agonist, an adrenalin
receptor antagonist, an adrenalin receptor agonist, an angiotensin
receptor agonist, a GABA receptor agonist, a thrombin receptor
antagonist, a thrombin receptor agonist, an opioid receptor
agonist, an ADP receptor agonist, a leukotriene receptor
antagonist, a leukotriene receptor agonist, a melatonin receptor
agonist, a somatostatin receptor agonist, a cannabinoid receptor
agonist, a sphingosine-1 phosphate receptor agonist, a metabotropic
glutamate receptor agonist, a phospholipase A2 inhibitor, a
TGF-.beta. production inhibitor, and a Th2 cytokine inhibitor, for
use as a cellular immunity induction promoter for transdermally
administering a WT1 peptide and/or a modified WT1 peptide; (57) A
method of inducing cellular immunity, comprising transdermally
administering to a subject (i) WT1 peptide and/or modified WT1
peptide and (ii) a pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof as a first cellular
immunity induction promoter; (58) A pharmacologically acceptable
acid or a pharmacologically acceptable salt thereof for use in
accelerating the induction of cellular immunity by the transdermal
administration of WT1 peptide and/or modified WT1 peptide; (59) A
combination of (i) a pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof and (ii) one or more
substances selected from the group consisting of TLR ligand, cyclic
dinucleotide, helper peptide, immunomodulatory small molecule drug,
cyclooxygenase inhibitor, prostaglandin receptor antagonist,
prostaglandin receptor agonist, TSLP production inhibitor,
adenylate cyclase inhibitor, omega-3 fatty acid, PPAR agonist,
dopamine receptor antagonist, dopamine receptor agonist, histamine
receptor agonist, histamine receptor antagonist, serotonin receptor
agonist, serotonin receptor antagonist, vasopressin receptor
antagonist, vasopressin receptor agonist, muscarine receptor
antagonist, muscarine receptor agonist, adrenalin receptor
antagonist, adrenalin receptor agonist, angiotensin receptor
agonist, GABA receptor agonist, thrombin receptor antagonist,
thrombin receptor agonist, opioid receptor agonist, ADP receptor
agonist, leukotriene receptor antagonist, leukotriene receptor
agonist, melatonin receptor agonist, somatostatin receptor agonist,
cannabinoid receptor agonist, sphingosine-1 phosphate receptor
agonist, metabotropic glutamate receptor agonist, phospholipase A2
inhibitor, TGF-beta production inhibitor and Th2 cytokine
inhibitor, for use in accelerating the induction of cellular
immunity by the transdermal administration of WT1 peptide and/or
modified WT1 peptide; (60) A combination of (i) WT1 peptide and/or
modified WT1 peptide and (ii) a pharmacologically acceptable acid
or a pharmacologically acceptable salt thereof as a first cellular
immunity induction promoter, for use in inducing cellular immunity
by the transdermal administration of WT1 peptide and/or modified
WT1 peptide; (61) A combination of (i) WT1 peptide and/or modified
WT1 peptide and (ii) a pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof for use in treating or
preventing a cancer, wherein the combination is transdermally
administered; and (62) Use of (i) WT1 peptide and/or modified WT1
peptide and (ii) a pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof as a first cellular
immunity induction promoter, for the manufacture of a cancer
vaccine composition for transdermal administration intended for the
induction of cellular immunity.
Since the cancer vaccine composition of the present invention can
be transdermally administered, it has the following advantages:
excellent compliance, for example, non-invasive administration, no
pain, and release from fear of injection; patients can administer
the cancer vaccine composition by himself/herself since the
administration is simple; a risk of accidental infection due to
needlestick injury by health care workers can be avoided; in the
case of repetitive administration, the ambulatory frequency can be
reduced, and this can contribute to the improvement in quality of
life of the patient; and medical wastes which necessitate special
disposition such as an injection needle are not generated. In
addition, if the cancer vaccine composition is in a form of a patch
preparation such as a cataplasm preparation or a tape preparation,
a predetermined dose can be surely administered, the drug releasing
rate can be arbitrarily controlled, and the cancer vaccine
composition is not adhered to other site upon administration.
Further, since the patch preparation can be easily detached, in the
case where a side effect is generated, the patient himself/herself
can instantaneously stop the administration by removing the patch
from the application site. Further, there is also an advantage that
efficacy of the cancer vaccine composition of the present invention
is remarkably improved, as compared with administration of the WT1
peptide and/or the modified WT1 peptide alone. Further, there is
also has an advantage that, by using a pharmacologically acceptable
acid or a pharmacologically acceptable salt thereof, efficacy of
the cancer vaccine composition for transdermal administration is
more improved. Further, the cancer vaccine composition of the
present invention also has an advantage that transdermal
administration of the composition induces stronger cellular
immunity as compared with injection administration.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a view showing cellular immunity inducing effect by a PIB
tape preparation with various acids incorporated therein.
FIG. 2 is a view showing the acid addition effect in various tape
preparations.
DETAILED DESCRIPTION OF THE INVENTION
First, terms used in the present specification will be defined so
that the present invention can be more easily understood. Terms
having no definition have the meaning which is normally understood
by a person skilled in the art in the fields of, particularly,
medicine, pharmacy, immunology, cell biology, biochemistry, polymer
chemistry and the like, unless the context requires otherwise.
I. Definition
As used herein, the term "WT1 peptide" means a partial peptide
consisting of about 8 to about 15, preferably about 8 to about 12
amino acids, which is obtained by fragmenting a WT1 protein which
is a product of a cancer gene WT1 (Wilm's tumor), and includes a
Db126 peptide, a Db235 peptide and the like. In addition, a partial
peptide of a WT1 product disclosed in WO 2000/06602, a WT1-derived
HLA-A26 binding cancer antigen peptide described in WO 2005/095598,
a HLA-A*3303-restricted WT1 peptide described in WO 2007/097358,
and a HLA-A*1101-restricted WT1 peptide described in WO 2008/081701
are also included in the "WT1 peptide" of the present
invention.
The term "Db126 peptide" means a WT1 peptide consisting of a
sequence Arg Met Phe Pro Asn Ala Pro Tyr Leu (SEQ ID No.: 1). The
term "Db235 peptide" means a WT1 peptide consisting of a sequence
Cys Met Thr Trp Asn Gln Met Asn Leu (SEQ ID No.: 2) (Patent
Document 1).
As used herein, the term "modified WT1 peptide" means a peptide in
which all or a part of amino acids of a WT1 peptide are modified by
substitution, modification or the like.
The modified WT1 peptide includes, for example,
(a) a peptide consisting of an amino acid sequence in which one to
several, for example, 1, 2, 3, 4 or 5 amino acids are substituted,
deleted or added in an amino acid sequence of a WT1 peptide;
and
(b) a peptide consisting of an amino acid sequence in which all or
a part of amino acids, for example, one or more, for example, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 amino acids are modified in an
amino acid sequence of a WT1 peptide.
Examples of "modification" of an amino acid which can be possessed
by a modified WT1 peptide include, but not limited to, aliphatic
chain addition modification such as alkylation such as acetylation
and methylation, glycosylation, hydroxylation, carboxylation,
aldehydization, phosphorylation, sulfonylation, formylation,
myristoylation, palmitoylation and stearoylation, octanoylation,
esterification, amidation, deamidation, disulfide bond formation
modification such as cystine modification, glutathione modification
and thioglycolic acid modification, glycation, ubiquitination,
succinimide formation, glutamylation, prenylation and the like. The
modified WT1 peptide may contain a combination of substitution,
deletion or addition of one or more amino acids, and modification
of one or more amino acids.
As a specific example, a Db235m peptide in which a part of a Db235
peptide is modified is a modified WT1 peptide consisting of a
sequence Cys Tyr Thr Trp Asn Gln Met Asn Leu (SEQ ID No.: 3) (WO
2002/079253), and is included in the modified WT1 peptide in the
present invention. A WT1 substitution type peptide described in WO
2004/026897, a WT1.sub.235-293 peptide derivative disclosed in WO
2007/063903 A1, and a HLA-A24 restrictive cancer antigen peptide
disclosed in WO 2003/106682 are also included in the modified WT1
peptide in the present invention. Specific examples of the HLA-A24
restrictive modified WT1 peptide described in WO 2003/106682
include a RYF peptide of a sequence Arg Tyr Phe Pro Asn Ala Pro Tyr
Leu (SEQ ID No.: 4), and an AYL peptide of a sequence Ala Tyr Leu
Pro Ala Val Pro Ser Leu (SEQ ID No.: 5).
The WT1 peptide and/or the modified WT1 peptide can be in the free
form or any pharmacologically acceptable salt form, for example, a
form of acid salts (acetic acid salt, TFA salt, hydrochloric acid
salt, sulfuric acid salt, phosphoric acid salt, lactic acid salt,
tartaric acid salt, maleic acid salt, fumaric acid salt, oxalic
acid salt, hydrobromic acid salt, succinic acid salt, nitric acid
salt, malic acid salt, citric acid salt, oleic acid salt, palmitic
acid salt, propionic acid salt, formic acid salt, benzoic acid
salt, picric acid salt, benzenesulfonic acid salt, dodecylsulfuric
acid salt, methanesulfonic acid salt, p-toluenesulfonic acid salt,
glutaric acid salt, various amino acid salts, etc.), metal salts
(alkali metal salts (e.g. sodium salt, potassium salt), alkaline
earth metal salts (e.g. calcium salt, magnesium salt), aluminum
salt etc.), or amine salts (triethylamine salt, benzylamine salt,
diethanolamine salt, t-butylamine salt, dicyclohexylamine salt,
arginine salt, dimethylammonium salt, ammonium salt, etc.). A
preferable pharmacologically acceptable salt is an acetic acid salt
or a TFA salt. The WT1 peptide and/or the modified WT1 peptide
which has been synthesized or produced, and isolated and purified
by a well-known method can be used.
As used herein, the term "cellular immunity induction promoter"
means any substance which can enhance the cellular immune response
induced by an antigen which is administered together with the
substance, as compared with the immune response induced by the
antigen without the substance. The cellular immunity induction
promoter may include substances specified in the present
specification, though it is not limited by the action mechanism by
which induction of the cellular immunity is promoted.
As used herein, the term "TLR ligand" means a ligand of a Toll-like
receptor (TLR), and includes, for example, ligands of TLR1 to 9.
Examples of the TLR ligand include a TLR1/2 ligand, a TLR2/6
ligand, a TLR2 and Dectin1 ligand, a TLR3 ligand, a TLR4 ligand, a
TLR5 ligand, a TLR7 and/or TLR8 ligand, a TLR9 ligand and the like.
In a preferable aspect of the present invention, the TLR ligand is
a TLR1/2 ligand, a TLR2 and Dectin1 ligand, a TLR3 ligand, a TLR4
ligand, a TLR7 and/or TLR8 ligand, and/or a TLR9 ligand.
As used herein, the term "TLR1/2 ligand" means a ligand of a
heterodimer of a Toll-like receptor (TLR) 1 and a Toll-like
receptor (TLR) 2, and includes, for example, a triacylated
lipoprotein derived from a cell wall of a bacterium and a salt
thereof, and these may be an extract, a product or a synthetic
product, and are not limited to them.
In a preferable aspect of the present invention, the TLR1/2 ligand
is Pam.sub.3CSK.sub.4. Pam.sub.3CSK.sub.4 has the formula
("Ser-Lys-Lys-Lys-Lys" disclosed as SEQ ID NO: 12):
##STR00001##
As used herein, the term "TLR2 and Dectin1 ligand" means a ligand
of a Toll-like receptor (TLR) 2 and a .beta.1,3-glucan receptor
(Dectin1), and includes, for example, a .beta.1,3-glucan derived
from a cell wall of a fungus and a salt thereof, and these may be
an extract, a product or a synthetic product, and are not limited
to them. In a preferable aspect of the present invention, the TLR2
and Dectin1 ligand is Zymosan derived from a yeast cell wall.
As used herein, the term "TLR3 ligand" means a ligand of a
Toll-like receptor (TLR) 3, and includes, for example, a
double-stranded RNA (dsRNA) derived from a virus and a salt
thereof, and these may be an extract, a product or a synthetic
product, and are not limited to them. In a preferable aspect of the
present invention, the TLR3 ligand is polyinosinic-polycytidylic
acid (Poly(I:C)) which is a synthetic product and/or a salt
thereof.
As used herein, the term "TLR4 ligand" means a ligand of a
Toll-like receptor (TLR) 4, and includes, for example, a
lipopolysaccharide (LPS) derived from a bacterium or a plant,
particularly, a lipid A derivative, for example, monophosphoryl
lipid A, a 3 deacylated monophosphoryl lipid A (3D-MPL), OM174, OM
294 DP or OM 197 MP-Ac DP and the like, alkyl glucosaminide
phosphate (AGP), for example, AGP disclosed in WO 98/50399 or U.S.
Pat. No. 6,303,347, or a salt of AGP as disclosed in U.S. Pat. No.
6,764,840, and a lipopolysaccharide, a glucopyranosyl lipid, and
sodium hyaluronate derived from a Pantoea bacterium, but is not
limited to them.
In a preferable aspect of the present invention, as the TLR4
ligand, lipopolysaccharides derived from genus Acetobacter (e.g.
Acetobacter aceti, Acetobacter xylinum, Acetobacter orientalis
etc.), genus Zymomonas (e.g. Zymomonas mobilis etc.), genus
Xanthomonas (e.g. Xanthomonas campestris etc.), genus Enterobacter
(e.g. Enterobacter cloacae etc.), and genus Pantoea (e.g. Pantoea
agglomerans etc.) are preferable. Extracts derived from these
lipopolysaccharides or purified lipopolysaccharides can be used as
they are. In addition, for example, lipopolysaccharides (IP-PA1)
derived from Pantoea agglomerans can be purchased from Funakoshi
Corporation. In addition, in a preferable aspect of the present
invention, the TLR4 ligand is a lipopolysaccharide, glucopyranosyl
lipid, and/or sodium hyaluronate derived from a Pantoea
bacterium.
As used herein, the term "TLR7 and/or TLR8 ligand" means a ligand
of a Toll-like receptor (TLR) 7 and/or TLR8, and includes, for
example, a single-stranded RNA, imiquimod, resiquimod (R848),
TLR7-II and other compounds, for example, loxoribine and
bropirimine, but is not limited to them.
In a preferable aspect of the present invention, the TLR7 and/or
TLR8 ligand is imiquimod. Imiquimod is
1-(2-methylpropyl)-1H-imidazo[4,5-c]quinoline-4-amine of the
formula:
##STR00002## and, for example, the characteristics and a production
process thereof are described in JP 7-505883 A (Patent Document
2).
In other preferable aspect, the TLR7 and/or TLR8 ligand is
resiquimod. Resiquimod is
4-amino-2-(ethoxymethyl)-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinol-
ine-1-ethanol of the formula:
##STR00003##
In other preferable aspect, the TLR7 and/or TLR8 ligand is TLR7-II.
TLR7-II is represented by the formula:
##STR00004##
In other preferable aspect, the TLR7 and/or TLR8 ligand is
bropirimine. Bropirimine is represented by the formula:
##STR00005##
As used herein, the term "TLR9 ligand" means a ligand of a
Toll-like receptor (TLR) 9, and includes, for example, ODN1826 and
the like. The TLR9 ligand used in the present invention may be an
extract, a product or a synthetic product, and is not limited to
them. In a preferable aspect of the present invention, the TLR9
ligand is ODN1826.
ODN1826 is an oligodeoxynucleotide consisting of the following
sequence (SEQ ID No.: 6).
TABLE-US-00001 5'-tccatgacgttcctgacgtt-3'
As used herein, the term "TLR2/6 ligand" means a ligand of a
heterodimer of Toll-like receptor (TLR) 2 and a Toll-like receptor
(TLR) 6, and includes, for example, a diacylated lipoprotein
derived from a cell wall of mycoplasma and a salt thereof, and
these may be an extract, a product or a synthetic product, and are
not limited to them. In a preferable aspect of the present
invention, the TLR2/6 ligand is Pam.sub.2CSK.sub.4, MALP-2 and/or
FSL-1.
Pam.sub.2CSK.sub.4 is represented by the following formula
("Ser-Lys-Lys-Lys-Lys" disclosed as SEQ ID NO: 12).
##STR00006##
FSL-1 is represented by the following formula
("Gly-Asp-Pro-Lys-His-Pro-Lys-Ser-Phe" disclosed as SEQ ID NO:
13).
##STR00007##
As used herein, the term "TLR5 ligand" means a ligand of a
Toll-like receptor (TLR) 5, and includes, for example, flagellin
and the like. The TLR5 ligand used in the present invention may be
an extract, a product or a synthetic product, and is not limited to
them. In a preferable aspect of the present invention, the TLR5
ligand is flagellin.
The Toll-like receptor (TLR) is a family of 1-type transmembrane
proteins which initiates congenital immune response in which a
specific cytokine, a specific chemokine and a growth factor
participate, by in vivo activation thereof. All TLRs can activate a
certain intracellular signal transmission molecule, for example, a
nuclearity factor .kappa.B (NF-.kappa.B) and a mitogen-activated
protein kinase (MAP kinase) or the like, while a specific
population of a cytokine and a chemokine which are released seems
to be inherent to each TLR. TLR3, 7, 8 and 9 include a subfamily of
TLR which is present in an endosome fraction or a lysosome fraction
of an immune cell (e.g. dendritic cell and monocyte). Specifically,
TLR3 is expressed by a wide range of cells such as a dendritic cell
and a fibroblast, TLR7 is expressed by a plasma cell-like dendritic
cell, and is expressed by a monocyte to a lesser extent, TLR8 is
expressed by a monocyte as well as a monocyte-derived dendritic
cell and a myelogenous dendritic cell, and TLR9 is expressed by a
plasma cell-like dendritic cell. This subfamily mediates
recognition of a microorganism nucleic acid (single-stranded RNA,
double-stranded RNA, single-stranded DNA etc.). Agonists of TLR3,
TLR7 and/or TLR8, and TLR9 stimulate production of a variety of
inflammatory cytokines (including, for example, interleukin-6,
interleukin-12, TNF-.alpha., and interferon-.gamma.). Such agonists
also promote increase in expression of a costimulating molecule
(e.g. CD40, CD80, and CD86), a major histocompatibility complex
molecule, and a chemokine receptor. I-type interferons (IFN.alpha.
and IFN.beta.) are produced by a cell upon activation with TLR7
and/or TLR8 agonists.
As used herein, the term "cyclic dinucleotide" means a molecule in
which two OH groups of a sugar part of two nucleotides produce an
ester for each same phosphoric acid molecule, and thereby
nucleotides are cyclized, and an analog thereof, and includes, for
example, cyclic di-AMP(c-di-AMP), cyclic di-GMP (c-di-GMP),
c-dGpGp, c-dGpdGp, c-GpAp, c-GpCp, c-GpUp and the like, but is not
limited to them. The cyclic dinucleotide activates a dendritic cell
or a T cell. Further examples of the cyclic dinucleotide, use of
them as an adjuvant, and a process for producing them are described
in JP 2007-529531 A (Patent Document 3). In a preferable aspect of
the present invention, the cyclic dinucleotide is cyclic di-GMP
and/or cyclic di-AMP. The cyclic di-GMP has the formula:
##STR00008## and a process for synthesizing it is described in
Kawai et al., Nucleic Acids Research Suppl. 3: 103-4.
As used in the present specification, the term "helper peptide"
means any peptide which activates a helper T cell, and includes,
for example, tubercle bacillus-derived helper peptide, measles
virus-derived helper peptide, hepatitis B virus-derived helper
peptide, hepatitis C virus-derived helper peptide, Chlamydia
trachomatis-derived helper peptide, Plasmodium falciparum
sporozoite-derived helper peptide, keyhole limpet
haemocyanin-derived helper peptide, tetanus toxin-derived helper
peptide, pertussis toxin-derived helper peptide, diphtheria
toxin-derived helper peptide, cancer cell-derived helper peptide
(e.g. WT1_.sub.332-347 helper peptide (described in Japanese Patent
No. 4621142 "WT1-derived HLA-DR binding Antigen Peptide"),
hWT1.sub.35 helper peptide, hWT1.sub.86 helper peptide,
hWT1.sub.294 helper peptide (above three kinds are described in WO
2010/123065 "Cancer Antigen Helper Peptide"), IMA-MMP-001 helper
peptide, CEA-006 helper peptide, MMP-001 helper peptide, TGFBI-004
helper peptide, HER-2/neu (aa776-790) helper peptide, AE36 helper
peptide, AE37 helper peptide, MET-005 helper peptide, BIR-002
helper peptide etc.), and universal helper analog (e.g. PADRE). In
a preferable aspect of the present invention, the helper peptide
consists of 10 to 20 amino acids, preferably 12 to 19 amino acids,
more preferably 13 to 18 amino acids. In a preferable aspect of the
present invention, the helper peptide is Peptide-25, hWT1.sub.35,
PADRE, or WT1_.sub.332-347. Peptide-25 is a peptide of 15 amino
acids consisting of a sequence Phe Gln Asp Ala Tyr Asn Ala Ala Gly
Gly His Asn Ala Val Phe (SEQ ID No.: 7), corresponding to amino
acid residues 240 to 254 of Ag85B which is one of main proteins
secreted by human tubercle bacillus (Mycobacterium tuberculosis).
Further, hWT1.sub.35 is a peptide of 18 amino acids consisting of a
sequence Trp Ala Pro Val Leu Asp Phe Ala Pro Pro Gly Ala Ser Ala
Tyr Gly Ser Leu (shown as SEQ ID No.: 8 in the present
application), described in WO 2010/123065 "Cancer Antigen Helper
Peptide". PADRE is a peptide of 13 amino acids consisting of a
sequence D-Ala Lys cyclohexyl-Ala Val Ala Ala Trp Thr Leu Lys Ala
Ala D-Ala (shown as SEQ ID No.: 9 in the present application).
WT.sup.1_.sub.332-347 is a peptide of 16 amino acids consisting of
a sequence Lys Arg Tyr Phe Lys Leu Ser His Leu Gln Met His Ser Arg
Lys His (shown as SEQ ID No.: 10 in the present application),
described in Japanese Patent No. 4621142 "WT1-derived HLA-DR
binding Antigen Peptide".
In addition, in the present invention, in place of the
aforementioned helper peptides, or in combination therewith,
peptides in which all or a part of amino acids of the helper
peptides are modified by substitution, modification, or the like
(hereinafter, referred to as "modified helper peptide") can also be
used.
The modified helper peptide includes, for example,
(a) a peptide consisting of an amino acid sequence in which one to
several, for example, 1, 2, 3, 4 or 5 amino acids are substituted,
deleted or added, in an amino acid sequence of the original helper
peptide; and
(b) a peptide consisting of an amino acid sequence in which all or
a part of amino acids, for example, one or more, for example, 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 amino
acids are modified, in an amino acid sequence of the original
helper peptide.
One example of the modified helper peptide is Peptide-25B.
Peptide-25B is one example of modified Peptide-25, in which a part
of amino acids of Peptide-25 are modified in order to enhance the
immunostimulation effect, and is a peptide of 15 amino acids
consisting of a sequence Phe Gln Asp Ala Tyr Asn Ala Val His Ala
Ala His Ala Val Phe (SEQ ID No. 11).
Examples of the "modification" of an amino acid which can be
possessed by the modified helper peptide include, but are not
limited to, aliphatic chain addition modification such as
acetylation, alkylation such as methylation, glycosylation,
hydroxylation, carboxylation, aldehydization, phosphorylation,
sulfonylation, formylation, addition of fatty acid such as
myristoylation, palmitoylation and stearoylation, octanoylation,
esterification, amidation, deamidation, disulfide bond formation
modification such as cystine modification, glutathione modification
and thioglycolic acid modification, glycation, ubiquitination,
succinimide formation glutamylation, prenylation and the like. In
addition, the modified helper peptide may contain a combination of
substitution, deletion or addition of one or more amino acids, and
modification of one or more amino acids.
As used herein, the term "cyclooxygenase inhibitor" means a
substance which inhibits the function of cyclooxygenase (COX). This
is also referred to as "COX inhibitor" hereinafter. As COX
inhibitors, there are a COX inhibitor which selectively acts on
particular cyclooxygenase (e.g. COX-1 or COX-2), and a COX
inhibitor having no selectivity. Examples of COX inhibitors which
can be used in the present invention include etodolac, loxoprofen,
celecoxib, valdecoxib, parecoxib, lumiracoxib, meloxicam,
tenoxicam, diclofenac, mefenamic acid, tolfenamic acid, flufenamic
acid, meclofenamic acid, niflumic acid, benzydamine, indobufen,
triflusal, tolmetin, fenoprofen, tiaprofenic acid, felbinac,
nepafenac, amfenac, pravadoline, zaltoprofen, sulindac, nabumetone,
diflunisal, piroxicam, ibuprofen, naproxen, fenoprofen, aspirin,
methyl salicylate, salicylamide, salsalate, aloxiprin, tolmetin,
indomethacin, proglumetacine, acemetacin, flurbiprofen,
pranoprofen, acetaminophen, floctafenine, lornoxicam, tenoxicam,
tiaprofenic acid, oxaprozin, ketoprofen, dexketoprofen,
dexibuprofen, alminoprofen, ketorolac, mofezolac, phenylbutazone,
oxyphenylbutazone, ketophenylbutazone, feprazone, phenbutazone,
ethenzamide, tiaramide, tinoridine, epirizole, emorfazone and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof. In a preferable aspect of the present invention, the COX
inhibitor is etodolac and/or loxoprofen.
Loxoprofen is represented by the formula:
##STR00009##
As used herein, the term "prostaglandin receptor antagonist" means
a substance having the function of preventing prostaglandin from
acting on a receptor, and includes, for example, an EP2 receptor
antagonist, an EP4 receptor antagonist, a DP receptor antagonist,
and an IP receptor antagonist.
As used herein, the term "EP2 receptor antagonist" means a
substance having the function of preventing prostaglandin E2 from
acting on an EP2 receptor. Examples of the EP2 receptor antagonist
include AH6809 and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
AH6809 is represented by the formula:
##STR00010##
As used herein, the term "EP4 receptor antagonist" means a
substance having the function of preventing prostaglandin E.sub.2
from acting on an EP4 receptor. Examples of the EP4 receptor
antagonist include GW627368X and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
GW627368X is represented by the formula:
##STR00011##
As used herein, the term "DP receptor antagonist" means a substance
having the function of preventing prostaglandin D.sub.2 from acting
on a DP receptor. Examples of the DP receptor antagonist include
S-5751, BWA868C and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
BWA868C is represented by the formula:
##STR00012##
As used herein, the term "IP receptor antagonist" means a substance
having the function of preventing prostaglandin I.sub.2 from acting
on an IP receptor. Examples of the IP receptor antagonist include
RO1138452 and a derivative thereof, as well as a pharmacologically
acceptable salt thereof.
RO1138452 is represented by the formula:
##STR00013##
As used herein, the term "prostaglandin receptor agonist" means a
substance having the function of acting on a prostaglandin
receptor, and includes, for example, an EP3 receptor agonist.
As used herein, the term "EP3 receptor agonist" means a substance
having the function of acting on an EP3 receptor. Examples of the
EP3 receptor agonist include sulprostone, GR63799, cloprostenol,
ONO-AE-248, carbacyclin, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Sulprostone is represented by the formula:
##STR00014##
As used herein, the term "TSLP production inhibitor" means a
substance having the function of inhibiting production of TSLP.
Since a drug which inhibits NF-.kappa.B is thought to indirectly
inhibit the production of TSLP, it is included in this category.
Examples of the TSLP production inhibitor include naringenin,
berberine, resveratrol, luteolin, apigenin, chrysoeriol, velutin,
rutin, hesperidin, quercetin, daidzein, genistein, noscapine,
diindolylmethane, xanthone, parthenolide and a derivative thereof,
as well as a pharmacologically acceptable salt thereof.
Berberine is represented by the formula:
##STR00015##
As used herein, the term "adenylate cyclase inhibitor" means a
substance having the function of inhibiting the activity of
adenylate cyclase. Examples of the adenylate cyclase inhibitor
include 2',5'-dideoxyadenosine, niacin, insulin, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
2',5'-Dideoxyadenosine is represented by the formula:
##STR00016##
As used herein, the term "omega-3 fatty acid" refers to an
unsaturated fatty acid having a carbon-carbon double bond at a
.omega.-3 position. Examples of the omega-3 fatty acid include
eicosapentaenoic acid, .alpha.-linolenic acid, docosahexaenoic
acid, and a derivative thereof, as well as a pharmacologically
acceptable salt thereof.
Eicosapentaenoic acid is represented by the formula:
##STR00017##
As used herein, the term "PPAR agonist" means a substance having
the function of acting on a peroxisome proliferator-activated
receptor, and includes, for example, a PPAR-.alpha. agonist, a
PPAR-.delta. agonist, and a PPAR-.gamma. agonist.
As used herein, the term "PPAR-.alpha. agonist" means a substance
having the function of acting on an .alpha. type peroxisome
proliferator-activated receptor. The term "PPAR-.delta. agonist"
means a substance having the function of acting on a .delta. type
peroxisome proliferator-activated receptor. The term "PPAR-.gamma.
agonist" means a substance having the function of acting on a
.gamma. type peroxisome proliferator-activated receptor. Examples
of the PPAR-.alpha. agonist, and/or the PPAR-.delta. agonist,
and/or the PPAR-.gamma. agonist include clofibrate, fenofibrate,
bezafibrate, ciprofibrate, etofibrate, telmisartan, oleyl
ethanolamide, tetradecylthioacetic acid, troglitazone,
pioglitazone, rosiglitazone, balaglitazone, rivoglitazone,
ciglitazone, darglitazone, edaglitazone, netoglitazone,
indeglitazar, tesaglitazar, muraglitazar, aleglitazar, and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Clofibrate is represented by the formula:
##STR00018##
As used herein, the term "dopamine receptor antagonist" means a
substance having the function of preventing dopamine from acting on
a receptor, and includes, for example, a D1 receptor antagonist,
and a D5 receptor antagonist.
As used herein, the term "D1 receptor antagonist" means a substance
having the function of preventing dopamine from acting on a D1
receptor. Examples of the D1 receptor antagonist include
benzazepine, fenoldopam, lorcaserin, SCH23390, SCH39166, LE300 and
a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
Benzazepine is represented by the formula:
##STR00019##
As used herein, the term "D5 receptor antagonist" means a substance
having the function of preventing dopamine from acting on a D5
receptor. Examples of the D5 receptor antagonist include SCH39166
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
SCH39166 is represented by the formula:
##STR00020##
As used herein, the term "dopamine receptor agonist" means a
substance having the function of acting on a dopamine receptor, and
includes, for example, a D2 receptor agonist, a D3 receptor
agonist, and a D4 receptor agonist.
As used herein, the term "D2 receptor agonist" means a substance
having the function of acting on a D2 receptor. Examples of the D2
receptor agonist include cabergoline, bromocriptine, pergolide,
ropinirole, talipexole, aripiprazole, lurasidone, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Ropinirole is represented by the formula:
##STR00021##
As used herein, the term "D3 receptor agonist" means a substance
having the function of acting on a D3 receptor. Examples of the D3
receptor agonist include piribedil, rotigotine, PD1289077, OH-DPAT
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
Rotigotine is represented by the formula:
##STR00022##
As used herein, the term "D4 receptor agonist" means a substance
having the function of acting on a D4 receptor. Examples of the D4
receptor agonist include flibanserin, ABT724, PD168077, CP226269
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
Flibanserin is represented by the formula:
##STR00023##
As used herein, the term "histamine receptor antagonist" means a
substance having the function of preventing histamine from acting
on a receptor, and includes, for example, a H1 receptor antagonist,
and a H2 receptor antagonist.
As used herein, the term "H1 receptor antagonist" means a substance
having the function of preventing histamine from acting on a H1
receptor. Examples of the H1 receptor antagonist include
ketanserin, thonzylamine, mepyramine, tripelenamine, dimethindene,
clemastine, bamipine, isothipendyl, chlorphenoxamine,
dimetotiazine, chlorpromazine, hydroxyzine, opipramol, betahistine,
cinnarizine, levocabastine, antazoline, diphenylpyraline,
carbinoxamine, doxylamine, alimemazine, cyclizine, meclozine,
levocetirizine, cyproheptadine, phenindamine, triprolidine,
azatadine, astemizole, terfenadine, acrivastine, ebastine,
desloratadine, rupatadine, bilastine, mizolastine, noberastine,
rocastine, temelastine, bepotastine, diphenhydramine,
chlorpheniramine, ketotifen, promethazine, cyproheptadine,
epinastine, olopatadine, bepotastine, astemizole, emedastine,
mequitazine, oxatomide, loratadine, fexofenadine, cetirizine,
azelastine, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Diphenhydramine is represented by the formula:
##STR00024##
As used herein, the term "H2 receptor antagonist" means a substance
having the function of preventing histamine from acting on a H2
receptor. Examples of the H2 receptor antagonist include
cimetidine, ranitidine, famotidine, nizatidine, roxatidine,
lafutidine, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Famotidine is represented by the formula:
##STR00025##
As used herein, the term "histamine receptor agonist" means a
substance having the function of acting on a histamine receptor,
and includes, for example, a H1 receptor agonist, a H3 receptor
agonist, and a H4 receptor agonist.
As used herein, the term "H1 receptor agonist" means a substance
having the function of acting on a H1 receptor. Examples of the H1
receptor agonist include 2-pyridylethylamine, 2-thiazolylethylamine
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
2-Pyridylethylamine is represented by the formula:
##STR00026##
As used herein, the term "H3 receptor agonist" means a substance
having the function of acting on a H3 receptor. Examples of the H3
receptor agonist include imethridine, imetit, immepip,
.alpha.-methylhistamine, proxyfan, and a derivative thereof, as
well as a pharmacologically acceptable salt thereof.
Proxyfan is represented by the formula:
##STR00027##
As used herein, the term "H4 receptor agonist" means a substance
having the function of acting on a H4 receptor. Examples of the H4
receptor agonist include 4-methylhistamine, VUF8430, immepip and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
4-Methylhistamine is represented by the formula:
##STR00028##
As used herein, the term "serotonin receptor antagonist" means a
substance having the function of preventing serotonin from acting
on a receptor, and includes, for example, a 5-HT2 receptor
antagonist, a 5-HT4 receptor antagonist, a 5-HT6 receptor
antagonist, and a 5-HT7 receptor antagonist.
As used herein, the term "5-HT2 receptor antagonist" means a
substance having the function of preventing serotonin from acting
on a 5-HT2 receptor. Examples of the 5-HT2 receptor antagonist
include pizotifen, risperidone, olanzapine, quetiapine,
aripiprazole, blonanserin, clozapine, paliperidone, ritanserin,
yohimbine, mesulergine, agomelatine, cyclobenzaprine, sarpogrelate,
methysergide, ketanserin, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Olanzapine is represented by the formula:
##STR00029##
As used herein, the term "5-HT4 receptor antagonist" means a
substance having the function of preventing serotonin from acting
on a 5-HT4 receptor. Examples of the 5-HT4 receptor antagonist
include piboserod, GR113808, GR125487, RS39604, SB204070 and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Piboserod is represented by the formula:
##STR00030##
As used herein, the term "5-HT6 receptor antagonist" means a
substance having the function of preventing serotonin from acting
on a 5-HT6 receptor. Examples of the 5-HT6 receptor antagonist
include cerlapirdine, clozapine and a derivative thereof, as well
as a pharmacologically acceptable salt thereof.
Cerlapirdine is represented by the formula:
##STR00031##
As used herein, the term "5-HT7 receptor antagonist" means a
substance having the function of preventing serotonin from acting
on a 5-HT7 receptor. Examples of the 5-HT7 receptor antagonist
include lurasidone, metergoline, and a derivative thereof, as well
as a pharmacologically acceptable salt thereof.
Metergoline is represented by the formula:
##STR00032##
As used herein, the term "serotonin receptor agonist" means a
substance having the function of acting on a serotonin receptor,
and includes, for example, a 5-HT1 receptor agonist, and a 5-HT2
receptor agonist.
As used herein, the term "5-HT1 receptor agonist" means a substance
having the function of acting on a 5-HT1 receptor. Examples of the
5-HT1 receptor agonist include piclozotan, tandospirone,
sumatriptan, zolmitriptan, eletriptan, rizatriptan, naratriptan,
almotriptan, frovatriptan, avitriptan, ergotamine, ergot alkaloid,
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
Zolmitriptan is represented by the formula:
##STR00033##
As used herein, the term "5-HT2 receptor agonist" means a substance
having the function of acting on a 5-HT2 receptor. Examples of the
5-HT2 receptor agonist include .alpha.-methyl-5-HT, agomelatine,
norfenfluramine, meta-chlorophenylpiperazine and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Agomelatine is represented by the formula:
##STR00034##
As used herein, the term "vasopressin receptor antagonist" means a
substance having the function of preventing vasopressin from acting
on a receptor, and includes, for example, a V2 receptor
antagonist.
As used herein, the term "V2 receptor antagonist" means a substance
having the function of preventing vasopressin from acting on a V2
receptor. Examples of the V2 receptor antagonist include tolvaptan,
mozavaptan, conivaptan, lixivaptan, and a derivative thereof, as
well as a pharmacologically acceptable salt thereof.
Mozavaptan is represented by the formula:
##STR00035##
As used herein, the term "vasopressin receptor agonist" means a
substance having the function of acting on a vasopressin receptor,
and includes, for example, a V1 receptor agonist.
As used herein, the term "V1 receptor agonist" means a substance
having the function of acting on a V1 receptor. Examples of the V1
receptor agonist include vasopressin, felypressin, desmopressin,
lypressin, terlipressin, ornipressin, argipressin, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Desmopressin is represented by the formula:
##STR00036##
As used herein, the term "muscarine receptor antagonist" means a
substance having the function of acting on a muscarine receptor,
and includes, for example, a M1 receptor antagonist, a M3 receptor
antagonist, and a M5 receptor antagonist.
As used herein, the term "M1 receptor antagonist" means a substance
having the function of preventing acetylcholine from acting on a M1
receptor. The term "M3 receptor antagonist" means a substance
having the function of preventing acetylcholine from acting on a M3
receptor. The term "M5 receptor antagonist" means a substance
having the function of preventing acetylcholine from acting on a M5
receptor. Examples of the M1 receptor antagonist, and/or the M3
receptor antagonist, and/or the M5 receptor antagonist include
pirenzepine, atropine, trimebutine, piperidolate, oxybutynin,
tropicamide, propiverine, tolterodine, solifenacin, darifenacin,
imidafenacin, oxyphencyclimine, tiotropium bromide, esoxybutynin,
tiquizium, and a derivative thereof, as well as a pharmacologically
acceptable salt thereof.
Oxybutynin is represented by the formula:
##STR00037##
As used herein, the term "muscarine receptor agonist" means a
substance having the function of acting on a muscarine receptor,
and includes, for example, a M1 receptor agonist, a M2 receptor
agonist, a M3 receptor agonist, a M4 receptor agonist, and a M5
receptor agonist.
As used herein, the term "M1 receptor agonist" means a substance
having the function of acting on a M1 receptor. The term "M2
receptor agonist" means a substance having the function of acting
on a M2 receptor. The term "M3 receptor agonist" means a substance
having the function of acting on a M3 receptor. The term "M4
receptor agonist" means a substance having the function of acting
on a M4 receptor. The term "M5 receptor agonist" means a substance
having the function of acting on a M5 receptor. Examples of the M1
receptor agonist, and/or the M2 receptor agonist, and/or the M3
receptor agonist, and/or the M4 receptor agonist, and/or the M5
receptor agonist include acetylcholine, aceclidine, alvameline,
talsaclidine, xanomeline, pilocarpine, cevimeline, bethanechol,
mazaticol, muscarine, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Bethanechol is represented by the formula:
##STR00038##
As used herein, the term "adrenalin receptor antagonist" means a
substance having the function of preventing adrenalin from acting
on a receptor, and includes, for example, an .alpha.1 receptor
antagonist, a .beta.1 receptor antagonist, a .beta.2 receptor
antagonist, and a .beta.3 receptor antagonist.
As used herein, the term ".alpha.1 receptor antagonist" means a
substance having the function of preventing adrenalin from acting
on an .alpha.1 receptor. Examples of the .alpha.1 receptor
antagonist include prazosin, doxazosin, bunazosin, trimazosin,
alfuzosin, silodosin, terazosin, tamusulosin, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Tamusulosin is represented by the formula:
##STR00039##
As used herein, the term ".beta.1 receptor antagonist" means a
substance having the function of preventing adrenalin from acting
on a .beta.1 receptor. The term ".beta.2 receptor antagonist" means
a substance having the function of preventing adrenalin from acting
on a .beta.32 receptor. The term ".beta.3 receptor antagonist"
means a substance having the function of preventing adrenalin from
acting on a .beta.3 receptor. Examples of the .beta.1 receptor
antagonist, and/or the .beta.2 receptor antagonist, and/or the
.beta.3 receptor antagonist include bopindolol, pindolol, timolol,
dichloroisoprenaline, alprenolol, carteolol, indenolol, bunitrolol,
penbutolol, propranolol, nadolol, nipradilol, tilisolol,
acebutolol, celiprolol, metoprolol, atenolol, bisoprolol,
betaxolol, practolol, bevantolol, butoxamine, carvedilol,
amosulalol, arotinolol, labetalol, and a derivative thereof, as
well as a pharmacologically acceptable salt thereof.
Propranolol is represented by the formula:
##STR00040##
As used herein, the term "angiotensin receptor agonist" means a
substance having the function of acting on an angiotensin receptor,
and includes, for example, an AT2 receptor agonist.
As used herein, the term "adrenalin receptor agonist" means a
substance having the function of acting on an adrenalin receptor,
and includes, for example, an .alpha.1 receptor agonist, and an
.alpha.2 receptor agonist.
As used herein, the term ".alpha.1 receptor agonist" means a
substance having the function of acting on an .alpha.1 receptor.
The term ".alpha.2 receptor agonist" means a substance having the
function of acting on an .alpha.2 receptor. Examples of the
.alpha.1 receptor agonist, and/or the .alpha.2 receptor agonist
include norepinephrine, norfenefrine, etilefrine, naphazoline,
phenylephrine, midodrine, methoxamine, oxedrine, metaraminol,
arbutamine, ephedrine, oxymetazoline, tetryzoline, xylometazoline,
tramazoline, pseudoephidrene, dipivefrine, amidephrine,
methylephedrine, rilmenidine, brimonidine, medetomidine, xylazine,
tizanidine, guanfacine, methyldopa, guanabenz, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Xylazine is represented by the formula:
##STR00041##
As used herein, the term "angiotensin receptor agonist" means a
substance having the function of acting on an angiotensin receptor,
and includes, for example, an AT2 receptor agonist.
As used herein, the term "AT2 receptor agonist" means a substance
having the function of acting on an AT2 receptor. Examples of the
AT2 receptor agonist include novokinin, angiotensin and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Angiotensin is represented by the formula:
##STR00042##
As used herein, the term "GABA receptor agonist" means a substance
having the function of acting on a GABA receptor, and includes, for
example, a GABA.sub.B receptor agonist.
As used herein, the term "GABA.sub.B receptor agonist" means a
substance having the function of acting on a GABA.sub.B receptor.
Examples of the GABA.sub.B receptor agonist include baclofen,
.gamma.-aminobutyric acid, arbaclofen and a derivative thereof, as
well as a pharmacologically acceptable salt thereof.
Baclofen is represented by the formula:
##STR00043##
As used herein, the term "thrombin receptor antagonist" means a
substance having the function of preventing thrombin from acting on
a receptor, and includes, for example, a PAR-1 receptor
antagonist.
As used herein, the term "PAR-1 receptor antagonist" means a
substance having the function of preventing thrombin from acting on
a PAR-1 receptor. Examples of the PAR-1 receptor antagonist include
vorapaxar, atopaxar, FR171113, RWJ56110, dabigatran, dabigatran
etexilate, melagatran, ximelagatran, hirudin, hirulog, argatroban
and a derivative thereof, as well as a pharmacologically acceptable
salt thereof.
Vorapaxar is represented by the formula:
##STR00044##
As used herein, the term "thrombin receptor agonist" means a
substance having the function of acting on a thrombin receptor, and
includes, for example, a PAR-1 receptor agonist.
As used herein, the term "PAR-1 receptor agonist" means a substance
having the function of acting on a PAR-1 receptor. Examples of the
PAR-1 receptor agonist include TRAP-6, TRAP-14, NAT6-NH.sub.2 and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
TRAP-6 is represented by the formula:
##STR00045##
As used herein, the term "opioid receptor agonist" means a
substance having the function of acting on an opioid receptor.
Examples of the opioid receptor agonist include trimebutine,
alvimopan, morphine, oxycodone, dihydrocodeine, diamorphine,
pethidine, pentazocine, buprenorphine, butorphanol, nalbuphine,
tilidine, dezocine, meptazinol, tapentadol, naltrexone, methadone,
ethylmorphine, hydrocodone, acetyldihydrocodeine, nalorphine,
loperamide, remoxipride, opipramol, and a derivative thereof, as
well as a pharmacologically acceptable salt thereof.
Buprenorphine is represented by the formula:
##STR00046##
As used herein, the term "leukotriene receptor antagonist" means a
substance having the function of preventing leukotriene from acting
on a receptor, and includes, for example, a CysLT1 receptor
antagonist, and a CysLT2 receptor antagonist.
As used herein, the term "CysLT1 receptor antagonist" means a
substance having the function of preventing leukotriene from acting
on a CysLT1 receptor. The term "CysLT2 receptor antagonist" means a
substance having the function of preventing leukotriene from acting
on a CysLT2 receptor. Examples of the CysLT1 receptor antagonist,
and/or the CysLT2 receptor antagonist include montelukast,
zafirlukast, pranlukast, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof. Examples of the
pharmacologically acceptable salt of montelukast include
montelukast sodium and the like.
Montelukast sodium is represented by the formula:
##STR00047##
As used herein, the term "leukotriene receptor agonist" means a
substance having the function of acting on a leukotriene receptor,
and includes, for example, a BLT receptor agonist.
As used herein, the term "BLT receptor agonist" means a substance
having the function of acting on a BLT receptor. Examples of the
BLT receptor agonist include leukotriene B4, CAY10583 and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Leukotriene B4 is represented by the formula:
##STR00048##
As used herein, the term "ADP receptor agonist" means a substance
having the function of acting on an ADP receptor. Examples of the
ADP receptor agonist include adenosine diphosphate, and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Adenosine diphosphate is represented by the formula:
##STR00049##
As used herein, the term "melatonin receptor agonist" means a
substance having the function of acting on a melatonin receptor.
Examples of the melatonin receptor agonist include melatonin,
perlapine, tasimelteon, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Melatonin is represented by the formula:
##STR00050##
As used in the present specification, the term "somatostatin
receptor agonist" means a substance having the function of acting
on a somatostatin receptor. Examples of the somatostatin receptor
agonist include somatostatin, somatostatin-14, octreotide, and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Octreotide is represented by the formula:
##STR00051##
As used herein, the term "cannabinoid receptor agonist" means a
substance having the function of acting on a cannabinoid receptor.
Examples of the cannabinoid receptor agonist include dronabinol,
nabilone, levonantradol, otenabant, GW833972A, GW405833, and a
derivative thereof, as well as a pharmacologically acceptable salt
thereof.
Dronabinol is represented by the formula:
##STR00052##
As used herein, the term "sphingosine-1 phosphate receptor agonist"
means a substance having the function of acting on a sphingosine-1
phosphate receptor. Examples of the sphingosine-1 phosphate
receptor agonist include fingolimod, ponesimod, RPC-1063, ONO-4641,
SEW2871, sphingosine-1 phosphate and a derivative thereof, as well
as a pharmacologically acceptable salt thereof.
Fingolimod is represented by the formula:
##STR00053##
As used herein, the term "metabotropic glutamate receptor agonist"
means a substance having the function of acting on a metabotropic
glutamate receptor, and includes, for example, an mGluR2 receptor
agonist, an mGluR3 receptor agonist, an mGluR4 receptor agonist, an
mGluR6 receptor agonist, an mGluR7 receptor agonist, and an mGluR8
receptor agonist.
As used herein, the term "mGluR2 receptor agonist" means a
substance having the function of acting on an mGluR2 receptor. The
term "mGluR3 receptor agonist" means a substance having the
function of acting on an mGluR3 receptor. The term "mGluR4 receptor
agonist" means a substance having the function of acting on an
mGluR4 receptor. The term "mGluR6 receptor agonist" means a
substance having the function of acting on an mGluR6 receptor. The
term "mGluR7 receptor agonist" means a substance having the
function of acting on an mGluR7 receptor. The term "mGluR8 receptor
agonist" means a substance having the function of acting on an
mGluR8 receptor. Examples of the mGluR2 receptor agonist, and/or
the mGluR3 receptor agonist, and/or the mGluR4 receptor agonist,
and/or the mGluR6 receptor agonist, and/or the mGluR7 receptor
agonist, and/or the mGluR8 receptor agonist include VU0361737,
VU0155041, biphenylindanone A, PBDA, L-AP4, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
VU0361737 is represented by the formula:
##STR00054##
As used herein, the term "phospholipase A2 inhibitor" means a
substance having the function of inhibiting the activity of
phospholipase A2. Examples of the phospholipase A2 inhibitor
include glycyrrhizic acid, glycyrrhetic acid, and a derivative
thereof, as well as a pharmacologically acceptable salt
thereof.
Glycyrrhetic acid is represented by the formula:
##STR00055##
As used herein, the term "TGF-.beta. production inhibitor" means a
substance having the function of inhibiting production of
TGF-.beta.. Examples of the TGF-.beta. production inhibitor include
pirfenidone, tranilast, and a derivative thereof, as well as a
pharmacologically acceptable salt thereof.
Pirfenidone is represented by the formula:
##STR00056##
As used herein, the term "Th2 cytokine inhibitor" means a substance
having the function of inhibiting production of a Th2 cytokine such
as IL-4 and IL-5. Examples of the Th2 cytokine inhibitor include
suplatast and a derivative thereof, as well as a pharmacologically
acceptable salt thereof. Examples of the pharmacologically
acceptable salt of suplatast include suplatast tosylate. In a
preferable aspect of the present invention, the Th2 cytokine
inhibitor is suplatast tosylate.
Suplatast tosylate is represented by the formula:
##STR00057##
As used herein, the term of "acid" means a Broensted acid, and
includes inorganic acids and organic acids, preferably carboxylic
acids, for example, fatty acid and lactic acid. As used in the
present specification, the "pharmacologically acceptable acid" as a
first cellular immunity induction promoter, which can be contained
in the composition of the present invention, means an acid which
has no harmful effect on an administration subject, and does not
lose the pharmacological activity of ingredients in the
composition. In a preferable aspect of the present invention, the
pharmacologically acceptable acid is an organic acid, more
preferably an organic compound containing carboxyl group or an
organic compound containing sulfonate group, more preferably
saturated or unsaturated straight or branched fatty acid in which a
saturated straight chain part has 8 to 20 carbon atoms, lactic
acid, malic acid, salicylic acid, maleic acid, citric acid, or an
organic compound containing sulfonate group, more preferably
saturated or unsaturated straight or branched fatty acid in which a
saturated straight chain part has 8 to 16 carbon atoms, lactic
acid, malic acid, salicylic acid, maleic acid, citric acid, or an
organic compound containing sulfonate group, further preferably
fatty acid selected from the group consisting of decanoic acid,
lauric acid, myristic acid, isostearic acid and oleic acid, or
lactic acid, salicylic acid, citric acid or methanesulfonic
acid.
As used herein, the "pharmacologically acceptable salt" which can
be contained in the composition of the present invention means a
salt which has no harmful effect on an administration subject, and
does not lose the pharmacological activity of ingredients in the
composition, and includes inorganic acid salts (e.g. hydrochloride
and phosphate), organic acid salts (e.g. acetate, phthalate, and
TFA salt), metal salts (alkali metal salts (e.g. sodium salt and
potassium salt), alkaline earth metal salts (e.g. calcium salt and
magnesium salt), aluminum salt etc.), amine salts (triethylamine
salt, benzylamine salt, diethanolamine salt, t-butylamine salt,
dicyclohexylamine salt, arginine salt, dimethylammonium salt,
ammonium salt etc.), but is not limited to them.
As used in the present specification, the term "immunomodulatory
small molecule drug" means a substance which activates or
suppresses immune cells such as a T cell, a NK cell, a macrophage
and the like, and which does not correspond to any of the
aforementioned TLR ligand, cyclic dinucleotide, helper peptide,
immunomodulatory small molecule drug, cyclooxygenase inhibitor,
prostaglandin receptor antagonist, prostaglandin receptor agonist,
TSLP production inhibitor, adenylate cyclase inhibitor, omega-3
fatty acid, PPAR agonist, dopamine receptor antagonist, dopamine
receptor agonist, histamine receptor agonist, histamine receptor
antagonist, serotonin receptor agonist, serotonin receptor
antagonist, vasopressin receptor antagonist, vasopressin receptor
agonist, muscarine receptor antagonist, muscarine receptor agonist,
adrenalin receptor antagonist, adrenalin receptor agonist,
angiotensin receptor agonist, GABA receptor agonist, thrombin
receptor antagonist, thrombin receptor agonist, opioid receptor
agonist, ADP receptor agonist, leukotriene receptor antagonist,
leukotriene receptor agonist, melatonin receptor agonist,
somatostatin receptor agonist, cannabinoid receptor agonist,
sphingosine-1 phosphate receptor agonist, metabotropic glutamate
receptor agonist, phospholipase A2 inhibitor, TGF-.beta. production
inhibitor, and Th2 cytokine inhibitor. Examples of the
immunomodulatory small molecule drug include bestatin, pidotimod,
levamisole, golotimod, forphenicinol, and a derivative thereof, as
well as a pharmacologically acceptable salt thereof. Examples of
the pharmacologically acceptable salt of levamisole include
levamisole hydrochloride and the like.
Bestatin is represented by the formula:
##STR00058##
Pidotimod is represented by the formula:
##STR00059##
Levamisole hydrochloride is represented by the formula:
##STR00060##
In the present invention, the immunomodulatory small molecule drug
is usually a compound having the molecular weight of less than
1000, preferably less than 500. In a preferable aspect of the
present invention, the immunomodulatory small molecule drug is one
or more compounds selected from the group consisting of bestatin,
pidotimod and levamisole hydrochloride.
As described above, the inventors have found that, among a variety
of cellular immunity induction promoters, a TLR ligand, a cyclic
dinucleotide, a helper peptide, an immunomodulatory small molecule
drug, a cyclooxygenase inhibitor, a prostaglandin receptor
antagonist, a prostaglandin receptor agonist, a TSLP production
inhibitor, an adenylate cyclase inhibitor, an omega-3 fatty acid, a
PPAR agonist, a dopamine receptor antagonist, a dopamine receptor
agonist, a histamine receptor agonist, a histamine receptor
antagonist, a serotonin receptor agonist, a serotonin receptor
antagonist, a vasopressin receptor antagonist, a vasopressin
receptor agonist, a muscarine receptor antagonist, a muscarine
receptor agonist, an adrenalin receptor antagonist, an adrenalin
receptor agonist, an angiotensin receptor agonist, a GABA receptor
agonist, a thrombin receptor antagonist, a thrombin receptor
agonist, an opioid receptor agonist, an ADP receptor agonist, a
leukotriene receptor antagonist, a leukotriene receptor agonist, a
melatonin receptor agonist, a somatostatin receptor agonist, a
cannabinoid receptor agonist, a sphingosine-1 phosphate receptor
agonist, a metabotropic glutamate receptor agonist, a phospholipase
A2 inhibitor, a TGF-.beta. production inhibitor, and a Th2 cytokine
inhibitor are particularly suitable for transdermally administering
the WT1 peptide antigen and/or the modified WT1 peptide antigen
and, therefore, in one aspect, a second cellular immunity induction
promoter which can be used together with a pharmacologically
acceptable acid or a pharmacologically acceptable salt thereof in
the present invention is selected from one or more kinds of them.
As a method of quantitatively measuring induction of cellular
immunity, a variety of methods have been developed, and one or more
of them, for example, the ELISPOT method described in Examples may
be used.
As used herein, the non-invasive administration means
administration without actively giving physical irritation and/or
chemical irritation, preferably without giving physical irritation
(e.g. without giving irritation by tape stripping or microneedle)
to a skin.
As used herein, the term "mildly irritating condition" means a
condition under which irritation to be given to the skin is lower
than the irritation generally given in order to improve the skin
permeability of the antigen contained in conventional vaccines, or
a condition under which irritation is not given to the skin at all.
In general, physical and/or chemical stimulation is given to the
skin before or simultaneously with the transdermal administration
of a conventional vaccine composition so that the antigen can
penetrate through the skin. In a preferable aspect of this
invention, examples of the mildly irritating condition include a
condition of low physical irritation and a condition of low
chemical irritation. The condition of low physical irritation is,
for example, a condition under which transepidermal water loss
(TEWL) (g/hm.sup.2) of a model animal for skin irritation
evaluation is 50 or less, preferably 45 or less, more preferably 40
or less, even more preferably 35 or less, still more preferably 30
or less. Since the TEWL level is about 2 (g/hm.sup.2) in the
non-treated skin, the TEWL level before the administration is 2
(g/hm.sup.2) or more. The condition of low chemical irritation is,
for example, a condition under which the thymic stromal
lymphopoietin (TSLP) level (pg/mg protein) in the skin of the model
animal for skin irritation evaluation is 10000 or less, preferably
9000 or less, more preferably 8000 or less, further preferably 7000
or less. Since the TSLP level is about 1 (pg/mg protein) in a
non-treated skin, the TSLP level at completion of the
administration of the vaccine composition exceeds 1 (pg/mg
protein), preferably exceeds 2 (pg/mg protein), more preferably
exceeds 3 (pg/mg protein). The "Thymic stromal lymphopoietin
(TSLP)" is a cytokine which participates in differentiation and
recruitment of a T cell, and can be utilized as an index of the
degree of skin irritation in the present invention. Greater TSLP
value means stronger skin irritation. Examples of means for
attaining the condition of low physical irritation include not
conducting the conventional pre-treatment of the skin before the
administration such as not conducting tape stripping, microneedle
puncture or the like before the administration. Examples of means
for attaining the condition of low chemical irritation include
avoiding administration of an irritating chemical ingredient such
as ethanol, a surfactant or the like at a certain amount or more.
The procedure for attaining the mildly irritating condition can be
determined by using a model animal for skin irritation evaluation,
and the determined procedure can be applied to the subject to be
treated by the vaccine composition, for example, a human
subject.
As used herein, the term "cancer" means a cancer associated with
abnormal expression, for example, overexpression of a WT1 gene.
Examples of cancer may include hematopoietic tumors or solid
cancers. Examples of the hematopoietic tumors associated with
abnormal expression of the WT1 gene include, but are not limited
to, leukemia such as acute myelocytic leukemia, acute lymphocytic
leukemia and chronic myelocytic leukemia, myelodysplastic syndrome,
multiple myeloma, as well as malignant lymphoma such as
non-Hodgkin's lymphoma. Examples of the solid cancers associated
with abnormal expression of the WT1 gene include, but are not
limited to, lung cancer, breast cancer, stomach cancer, large
intestine/rectum cancer, germ cell cancer, liver cancer, skin
cancer, pancreas cancer, bile duct cancer, head and neck squamous
cell cancer, thyroid cancer, kidney cancer, bladder cancer,
prostate cancer, ovarian cancer, uterine cancer, cervical cancer,
bone soft tissue sarcoma, malignant melanoma, malignant
mesothelioma, testicular germ cell tumor and malignant glioma.
As used herein, the term "abnormal expression of a gene" means that
the expression level of the gene in a cell is increased or
decreased remarkably, for example, by 2 times or more such as 4
times or more, as compared with the other cells in the same tissue.
The term "overexpression" means that the abnormal expression is an
increase in the expression level. The expression level of a gene
can be easily measured using any method well-known in the art.
As used herein, the term "subject" means any animal having the WT1
gene whose immune response can be induced by the transdermal
administration of a cancer vaccine composition for transdermal
administration at a practical stage. Typically the subject may be a
mammal such as a human, mouse, rat, dog, cat, rabbit, horse, cow,
sheep, pig, goat, monkey, chimpanzee or the like. A particularly
preferable subject is human.
As used herein, the term "model animal for immunological
evaluation" means a model animal for evaluating the property of a
cancer vaccine composition for transdermal administration immunity.
Specifically, it means a model animal for evaluating the property
of inducing cellular immunity. The model animal for immunological
evaluation should be delected in view of compatibility between the
antigen in the vaccine composition to be exalated and the MHC class
I molecule of the animal. An animal model suitable for evaluating
the property of the vaccine composition to induce cellular immunity
should be used. For example, in the case of a vaccine composition
comprising a HLA-A*24 type MHC restricted class I peptide, the
property may be evaluated by a BALB/c mouse. In the case of a
vaccine composition comprising a HLA-A*02 type MHC restricted
peptide, the property may be evaluated in a genetically modified
mouse by which immunity induction by the HLA-A*02 type MHC
restricted peptide can be evaluated. In the case of a vaccine
composition comprising other HLA type MHC restricted peptide, the
property is evaluated by an animal by which immunity induction by
the HLA type MHC restricted peptide can be evaluated. In the case
of a vaccine composition comprising a protein antigen, the property
is evaluated by an animal having MHC compatible with a class I
epitope to be used to induce immunity, among various class I
epitopes included in the an amino acid sequence of the protein
antigen. In addition, in the case of a cancer vaccine composition
for transdermal administration using Db126 peptide which is
compatible with not only HLA-A*02 type but also MHC-H-2Db type, not
only a genetically modified mouse by which immunity induction by
the HLA-A*0201 type MHC restricted peptide can be evaluated, but
also a C57BL/6 mouse which is an animal having MHC-H-2 Db type can
be used as the model animal for immunological evaluation. When the
hair of the animal is cut to ensure the place for transdermal
administration, the animal should be used after it is recovered
from the skin damage caused by the hair cut.
As used herein, the term "model animal for skin irritation
evaluation" means a model animal for evaluating transepidermal
water loss (TEWL) as an index of physical irritation of the skin,
or a model animal for evaluating TSLP as an index of the skin
irritation property of a cancer vaccine composition for transdermal
administration. Regardless of the kind of the antigen contained in
the cancer vaccine composition for transdermal administration,
C57BL/6 mouse may be used as model animal for skin irritation
evaluation. When the hair of the animal is cut to ensure the place
for the transdermal administration, the animal should be used after
it is recovered from the skin damage caused by the hair cut.
II. Cancer Vaccine Composition for Transdermal Administration
It has already been revealed that WT1 peptides and/or the modified
WT1 peptides are useful as a cancer vaccine (e.g. Patent Document
1).
As used herein, the term composition "for transdermal
administration" may be provided in any preparation or formulation
which is usually used for the transdermal administration, for
example, a liquid formulation for external use such as a liniment
formulation or a lotion formulation, a spray formulation for
external use such as an aerosol formulation, an ointment
formulation, a plaster formulation, a cream formulation, a gel
formulation, or a patch formulation such as a tape preparation or a
cataplasm formulation. Grouping, definition, nature, production
process and the like of these preparations or formulation are
well-known in the art. For example, see Japanese Pharmacopoeia
16.sup.th edition. A composition for transdermal administration
suitable in the present invention is in a form of a cream
formulation, a liquid formulation for external use or a tape
preparation.
Examples of the base for the liniment formulation include water;
alcohols such as ethanol and propylene glycol; fat oils such as
hard paraffin, soft paraffin, liquid paraffin, glycerin, paraffin
oil, beeswax and metal soap; mucilage; natural oils (e.g. almond
oil, corn oil, peanut oil, castor oil, olive oil, or a derivative
thereof (such as polyoxyl castor oil)); mutton tallow or a
derivative thereof, fatty acids and/or esters (e.g. stearic acid,
oleic acid, isopropyl myristate), as well as a mixture thereof.
The lotion formulation is a preparation in which the active
ingredient is finely and homogenously dispersed in an aqueous
liquid, and there are a suspending lotion formulation, and an
emulsified lotion formulation. Examples of the suspending agent
include gum arabic, sodium alginate, carboxymethylcellulose sodium,
methylcellulose, bentonite and the like. Examples of the
emulsifying agent include sodium lauryl sulfate, sorbitan fatty
acid ester and the like.
For example, as an ointment base, hydrophobic base such as fats or
oils, waxes and hydrocarbon compounds can be generally used.
Specifically, examples of the ointment base include mineral bases
such as yellow vaseline, white vaseline, paraffin, liquid paraffin,
plastibase, silicone and the like, and animal or plant bases such
as beeswax, animal or vegetable fat or oil and the like.
Examples of the base for cream formulation include water/oil type
bases such as hydrophilic ointment, vanishing cream and the like;
and oil/water type bases such as hydrophilic vaseline, purified
lanolin, Aquahole, Eucerin, Neocerin, hydrous lanolin, cold cream,
hydrophilic plastibase and the like.
As a gel base, for example, the followings can be used: a
carboxyvinyl polymer, a gel base, a fat-free ointment,
polyvinylpyrrolidone, polyvinyl alcohol, sodium polyacrylate,
carboxymethylcellulose, starch, xanthan gum, karaya gum, sodium
alginate, methylcellulose, hydroxypropylcellulose,
hydroxypropylmethylcellulose phthalate (HPMCP), cellulose acetate
phthalate (CAP), carboxymethylethylcellulose (CMEC),
ethylcellulose, hydroxyethylcellulose,
hydroxypropylmethylcellulose, a carboxyvinyl polymer, tragacanth,
gum arabic, tara gum, tamarind seed gum, psyllium seed gum, agar,
gellan gum, glucomannan, locust bean gum, guar gum, carrageenan,
dextrin, dextran, amylose, carboxymethylcellulose potassium,
carboxymethylcellulose sodium, carboxymethylcellulose calcium,
pullulan, chitosan, sodium carboxymethyl starch, plantago testa,
galactomannan, aminoalkyl methacrylate copolymer E, aminoalkyl
methacrylate copolymer RS, methacrylic acid copolymer L,
methacrylic acid copolymer LD, methacrylic acid copolymer S, methyl
acrylate/methacrylic acid/methyl methacrylate copolymer, ethyl
acrylate/methyl methacrylate copolymer, polyvinylacetal
diethylaminoacetate, casein, alginic acid alkyl ester, gelatin,
polyethylene glycol and the like as a hydrogel base.
Examples of the base for a cataplasm preparation include gelatin,
carboxymethylcellulose sodium, methylcellulose, sodium
polyacrylate, kaolin, polyvinyl alcohol, polyvinylpyrrolidone,
glycerin, propylene glycol, water and the like.
For example, a tape preparation comprises an adhesive layer
comprising an acrylic adhesive, a natural rubber adhesive, a
synthetic rubber adhesive (including rubber elastomer such as
synthetic isoprene rubber, polyisobutylene (PIB), styrene-butadiene
rubber, styrene-isoprene-styrene (SIS) rubber etc.), a silicone
adhesive, a vinyl ester adhesive, a vinyl ether adhesive or the
like, and a support which supports the adhesive layer. Optionally,
the preparation may further contain a release liner which covers
the adhesive layer to avoid exposure thereof before use and can be
easily peeled from the adhesive layer upon use.
The amount of the WT1 peptide and/or the modified WT1 peptide, the
pharmacologically acceptable acid or a pharmacologically acceptable
salt thereof, and the cellular immunity induction promoter in the
cancer vaccine composition for transdermal administration of the
present invention is not particularly limited. In one aspect, the
cancer vaccine composition for transdermal administration of the
present invention contains the WT1 peptide and/or the modified WT1
peptide preferably in an amount of 0.01 to 40% by weight, more
preferably 0.1 to 30% by weight based on the total weight of the
composition. In one aspect, the cancer vaccine composition for
transdermal administration of the present invention contains the
pharmacologically acceptable acid or a pharmacologically acceptable
salt thereof preferably in an amount of 0.001 to 30% by weight,
more preferably 0.01 to 20% by weight based on the total weight of
the composition. When the cancer vaccine composition for
transdermal administration of the present invention contains a
cellular immunity induction promoter, the cellular immunity
induction promoter is contained preferably in an amount of 0.001 to
30% by weight, more preferably 0.01 to 20% by weight based on the
total weigh of the composition.
When the cancer vaccine composition for transdermal administration
of the present invention is provided in the form of a tape
preparation, an adhesive layer of the tape preparation
(hereinafter, also referred to as "tape preparation of the present
invention") comprises an antigen and, optionally, further comprises
a cellular immunity induction promoter. In one aspect, the adhesive
layer of the tape preparation of the present invention comprises an
antigen preferably in an amount of 0.01 to 40% by weight, more
preferably 0.1 to 30% by weight based on the total weight of the
adhesive layer. When the adhesive layer of the tape preparation of
the present invention comprises a cellular immunity induction
promoter, the cellular immunity induction promoter is comprised
preferably in an amount of 0.001 to 30% by weight, more preferably
0.01 to 20% by weight based on the total weight of the adhesive
layer.
An adhesive which is to form the adhesive layer of the tape
preparation of the present invention is not particularly limited,
and examples thereof include acrylic adhesives consisting of an
acrylic polymer; rubber adhesives comprising a rubber elastomer
such as a styrene-diene-styrene block copolymer (e.g.
styrene-isoprene-styrene block copolymer, styrene-butadiene-styrene
block copolymer etc.), polyisoprene, polyisobutylene, butyl rubber,
polybutadiene and the like; silicone adhesives such as silicone
rubber, dimethylsiloxane base, diphenylsiloxane base and the like;
vinyl ether adhesives such as polyvinyl methyl ether, polyvinyl
ethyl ether, polyvinyl isobutyl ether and the like; vinyl ester
adhesives such as vinyl acetate-ethylene copolymer and the like;
and polyester adhesives consisting of a carboxylic acid component
such as dimethyl terephthalate, dimethyl isophthalate, dimethyl
phthalate, and a polyhydric alcohol component such as ethylene
glycol. A particularly preferable adhesive is an acrylic adhesive,
a rubber adhesive, and a silicone adhesive. These adhesives are
contained in the adhesive layer preferably in an amount of 10 to
90% by weight, more preferably 20 to 80% by weight, as a solid
matter thereof, based on the total weight of the adhesive
layer.
Examples of the acrylic adhesive include an acrylic acid ester
adhesive containing, as a main component, a polymer comprising
(meth)acrylic acid C2-C18 alkyl ester as a first monomer. Examples
of the (meth)acrylic acid alkyl ester (first monomer) include
(meth)acrylic acid alkyl esters in which an alkyl group is a
straight, branched or cyclic alkyl group having 1 to 18 carbon
atoms (e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl,
cyclohexyl, heptyl, octyl, 2-ethylhexyl, nonyl, decyl, undecyl,
dodecyl, tridecyl etc.). Preferred are (meth)acrylic acid alkyl
esters in which an alkyl group is a straight, branched or cyclic
alkyl group having 4 to 18 carbon atoms (e.g. butyl, pentyl, hexyl,
cyclohexyl, heptyl, octyl, 2-ethylhexyl, nonyl, decyl, undecyl,
dodecyl, tridecyl etc.). Further, since use of a monomer component
which lowers the glass transition temperature of a polymer is
suitable in order to impart adhesiveness at room temperature,
(meth)acrylic acid alkyl esters in which an alkyl group is a
straight, branched or cyclic alkyl group having 4 to 8 carbon atoms
(e.g. butyl, pentyl, hexyl, cyclohexyl, heptyl, octyl, 2-ethylhexyl
etc., preferably butyl, 2-ethylhexyl, and cyclohexyl, particularly
preferably 2-ethylhexyl) are more preferable. Specifically, butyl
acrylate, 2-ethylhexyl acrylate, 2-ethylhexyl methacrylate,
cyclohexyl acrylate, cyclohexyl methacrylate and the like are more
preferable and, among them, 2-ethylhexyl acrylate is most
preferable. These (meth)acrylic acid alkyl esters (first monomer
component) can be used alone, or can be used by combining two or
more thereof.
In addition, the acrylic adhesive may contain a second monomer
copolymerizable with the (meth)acrylic acid alkyl ester, and
examples of the second monomer include monomers having a functional
group which can become a crosslinking point upon use of a
crosslinking agent. Examples of the functional group which can
participate in a crosslinking reaction include a hydroxy group, a
carboxyl group, a vinyl group and the like, and a hydroxy group and
a carboxyl group are preferable. Specific examples of the monomer
(second monomer component) include (meth)acrylic acid hydroxyethyl
ester, (meth)acrylic acid hydroxypropyl ester,
N-hydroxyalkyl(meth)acrylamide, (meth)acrylic acid, itaconic acid,
maleic acid, maleic anhydride, mesaconic acid, citraconic acid,
glutaconic acid and the like. Among them, from the viewpoint of
easy availability, acrylic acid, methacrylic acid, and acrylic acid
hydroxyethyl ester (particularly, 2-hydroxyethyl acrylate) are
preferable, and acrylic acid is most preferable. These monomers
(second monomer component) can be used alone, or can be used by
combining two or more thereof.
Further, the acrylic adhesive may optionally contain a third
monomer in addition to the second monomer. Examples of the third
monomer (third monomer component) include vinyl esters such as
vinyl acetate, vinyl propionate and the like; vinyl ethers such as
methyl vinyl ether, ethyl vinyl ether and the like; vinyl amides
such as N-vinyl-2-pyrrolidone, N-vinylcaprolactam and the like;
(meth)acrylic acid alkoxy esters such as (meth)acrylic acid
methoxyethyl ester, (meth)acrylic acid ethoxyethyl ester,
(meth)acrylic acid tetrahydrofurfuryl ester and the like; hydroxy
group-containing monomers (since this is used as a third monomer
component, it is not a crosslinking point) such as hydroxypropyl
(meth)acrylate, .alpha.-hydroxymethyl acrylate and the like;
(meth)acrylic acid derivatives having an amide group such as
(meth)acrylamide, dimethyl(meth)acrylamide,
N-butyl(meth)acrylamide, N-methylol(meth)acrylamide and the like;
(meth)acrylic acid aminoalkyl esters such as (meth)acrylic acid
aminoethyl ester, (meth)acrylic acid dimethylaminoethyl ester,
(meth)acrylic acid t-butylaminoethyl ester and the like;
(meth)acrylic acid alkoxyalkylene glycol esters such as
(meth)acrylic acid methoxyethylene glycol ester, (meth)acrylic acid
methoxydiethylene glycol ester, (meth)acrylic acid
methoxypolyethylene glycol ester, (meth)acrylic acid
methoxypolypropylene glycol ester and the like;
(meth)acrylonitriles; monomers having sulfonic acid such as
styrenesulfonic acid, allylsulfonic acid,
sulfopropyl(meth)acrylate, (meth)acryloyloxynaphthalenesulfonic
acid, acrylamidemethylsulfonic acid and the like; and
vinyl-group-containing monomers such as vinylpiperidone,
vinylpyrimidine, vinylpiperazine, vinylpyrrole, vinylimidazole,
vinyloxazole, vinylmorpholine and the like. Among them, vinyl
esters, and vinyl amides are preferable, vinyl acetate is
preferable as vinyl esters, and N-vinyl-2-pyrrolidone is preferable
as vinyl amides. These monomers (third monomer component) can be
used alone, or can be used by combining two or more thereof.
When the acrylic adhesive is a copolymer of a (meth)acrylic acid
alkyl ester (first monomer component) and a vinyl monomer having a
functional group which can participate in a crosslinking reaction
(second monomer component), the (meth)acrylic acid alkyl ester and
the vinyl monomer having a functional group which can participate
in a crosslinking reaction are copolymerized by blending the
components at a weight ratio of (meth)acrylic acid alkyl
ester:vinyl monomer having a functional group which can participate
in a crosslinking reaction of preferably 99 to 85:1 to 15, more
preferably 99 to 90:1 to 10.
Further, when the acrylic adhesive is a copolymer of a
(meth)acrylic acid alkyl ester (first monomer component), a vinyl
monomer having a functional group which can participate in a
crosslinking reaction (second monomer component), and a monomer
other than them (third monomer component), the (meth)acrylic acid
alkyl ester, the vinyl monomer having a functional group which can
participate in a crosslinking reaction, and the monomer other than
them are copolymerized by blending the components at a weight ratio
of (meth)acrylic acid alkyl ester:vinyl monomer having a functional
group which can participate in a crosslinking reaction:monomer
other than them of preferably 40 to 94:1 to 15:5 to 50, more
preferably 50 to 89:1 to 10:10 to 40.
The components may be polymerized by a known method. For example,
the monomers in a solvent such as ethyl acetate may be reacted in
the presence of a polymerization initiator (e.g. benzoyl peroxide,
azobisisobutyronitrile etc.) at 50 to 70.degree. C., for 5 to 48
hours.
Particularly preferable acrylic adhesives in the present invention
are, for example, a copolymer of acrylic acid 2-ethylhexyl
ester/acrylic acid/N-vinyl-2-pyrrolidone, a copolymer of acrylic
acid 2-ethylhexyl
ester/N-(2-hydroxyethyl)acrylamide/N-vinyl-2-pyrrolidone, a
copolymer of acrylic acid 2-ethylhexyl ester/acrylic acid
2-hydroxyethyl ester/vinyl acetate and a copolymer of acrylic acid
2-ethylhexyl ester/acrylic acid and, more preferably, a copolymer
of acrylic acid 2-ethylhexyl ester/acrylic
acid/N-vinyl-2-pyrrolidone.
Optionally, these acrylic adhesives may be subjected to physical
crosslinking treatment by ultraviolet irradiation, or radiation
irradiation such as electron beam irradiation, or chemical
crosslinking treatment using various crosslinking agents such as an
isocyanate compound such as trifunctional isocyanate, organic
peroxide, organic metal salt, metal alcoholate, metal chelate
compound, polyfunctional compound (polyfunctional external
crosslinking agent, a monomer for polyfunctional internal
crosslinking such as diacrylate and dimethacrylate).
Examples of the rubber adhesive include rubber adhesives in which a
rubber elastomer such as polyisobutylene/polybutene elastomer,
styrene/diene/styrene block copolymer, styrene/butadiene elastomer,
nitrile elastomer, chloroprene elastomer, vinylpyridine elastomer,
polyisobutylene elastomer, butyl elastomer, or isoprene/isobutylene
elastomer is blended. Among them, in view of solubility of the
peptide and the cellular immunity induction promoter in the
adhesive and the skin adhesiveness, polyisobutylene (PIB),
styrene/diene/styrene block copolymer (e.g.
styrene/butadiene/styrene block copolymer (SBS),
styrene/isoprene/styrene block copolymer (SIS) etc.) and the like
are preferably used. A mixture of two or more of those adhesives
may also be used.
Further, in order to achieve a suitable adhesive force and drug
solubility of the rubber adhesive, the rubber adhesive may be a
mixture of two or more rubber elastomers of the same or different
monomer components each having different average molecular weights.
For example, with respect to polyisobutylene, a mixture of
polyisobutylene of high molecular weight having an average
molecular weight of 150,000 to 5,500,000, polyisobutylene of medium
molecular weight having an average molecular weight of 10,000 to
150,000 and/or polyisobutylene of low molecular weight having an
average molecular weight of 500 to 4,000 is preferable. In this
case, it is preferable to blend polyisobutylenes of high molecular
weight, medium molecular weight and low molecular weight at a
weight ratio of high molecular weight:medium molecular weight:low
molecular weight=10 to 80, preferably 20 to 70:0 to 90, preferably
10 to 80:0 to 80, preferably 10 to 60.
As used herein, the average molecular weight means the viscosity
average molecular weight calculated from the viscosity expression
of Flory, and is obtained by calculating the Staudinger index
(J.sub.0) from the flow time of the capillary 1 of a Ubbelohde
viscometer at 20.degree. C. by the Schulz-Blaschke expression, and
using this J.sub.0 value with the following expression.
J.sub.0=.eta..sub.sp/c(1+0.31.eta..sub.sp) (Schulz-Blaschke
equation) .eta..sub.sp=t/t.sub.0-1 (Formula) t: Flow time of
solution (according to Hagenbach-couette correction formula)
t.sub.0: Flow time of solvent (according to Hagenbach-couette
correction formula) c: Concentration of solution (g/cm.sup.3)
J.sub.0=3.06.times.10.sup.-2Mv.sup.0.65 Mv: Viscosity average
molecular weight
In order to impart suitable adhesiveness, for example, a tackifier
such as a rosin resin, a polyterpene resin, a coumarone-indene
resin, a petroleum resin, a terpene-phenol resin, a xylene resin,
an alicyclic saturated hydrocarbon resin or the like may be blended
in the rubber adhesive. One or two or more kinds of tackifiers can
be blended in an amount of 50% by weight or less, preferably 5 to
40% by weight based on the total weight of the rubber adhesive.
Examples of the silicone adhesive include silicone adhesives
consisting of polyorganosiloxane adhesive, polydimethylsiloxane
adhesive, polydimethyldiphenyl-siloxane adhesive. Inter alia, a
commercially available silicone adhesive such as BIO PSA from Dow
Corning Corporation is preferably used.
The support which supports the adhesive layer is not particularly
limited, and a support that is substantially impervious to the
peptide and the cellular immunity induction promoter so that the
peptide, the cellular immunity induction promoter, additives or the
like contained in the adhesive layer will not pass through the
support and leaked from the rear surface.
As the support, for example, a single film of polyester, polyamide,
poly(vinylidene chloride), polyethylene, polypropylene, poly(vinyl
chloride), ethylene-ethyl acrylate copolymer,
polytetrafluoroethylene, ionomer resin, metal foil or the like, or
a laminate film of them can be used. Among them, in order to make
adhesiveness (anchorability) between the support and the adhesive
layer good, it is preferable that the support is a laminate film of
a nonporous plastic film and a porous film made of the
aforementioned material. In this case, it is desirable that the
adhesive layer is formed on the porous film side. As such a porous
film, a porous film which improves anchorability with the adhesive
layer is adopted, and specific examples thereof include a paper
sheet, a woven fabric, a non-woven fabric, a knitted fabric, a
sheet which has been mechanically perforation-treated, and the
like. Among them, from the viewpoint of handling property and the
like, particularly, a paper sheet, a woven fabric and a non-woven
fabric are preferable. As the porous film, in view of improvement
in anchorability, softness and sticking operability of a tape
preparation and the like, a porous film having a thickness in the
range of 1 to 200 .mu.m is adopted. In addition, when a woven
fabric or a non-woven fabric is used as the porous film, the weight
per unit area is preferably 5 to 30 g/m.sup.2, more preferably 6 to
15 g/m.sup.2.
Examples of a most suitable support include a laminate film of a
polyester film (preferably, polyethylene terephthalate film) having
a thickness of 1.5 to 6 .mu.m, and a non-woven fabric made of
polyester (preferably, polyethylene terephthalate) having a weight
per unit area of 6 to 15 g/m.sup.2.
In the tape preparation of the present invention, in order to
protect the surface of the adhesive layer until use, it is
desirable that a release liner is laminated on the adhesive
surface. The release liner is not particularly limited as far as it
is treated so that it has the releasing property and it can be
released with a sufficiently small peeling force. For example, a
film of polyester, poly(vinyl chloride), poly(vinylidene chloride),
polyethylene terephthalate or the like, paper such as pure paper,
glassine paper and the like, or a laminate film of pure paper or
glassine paper and polyolefin may be treated with the adhesive
layer and is used as the release liner. The thickness of the
release liner is preferably 10 to 200 .mu.m, more preferably 25 to
100 .mu.m. As the release liner, polyester layer, particularly,
polyethylene terephthalate layer is preferable in view of the
barrier property and the cost. Further, in this case, in terms of
handling property, a release liner having a thickness of around 25
to 100 .mu.m is preferable.
It is preferable that the composition of the present invention is
administered to a subject under the mildly irritating condition.
Administration under the mildly irritating condition can be
attained by, for example, (i) administering the composition of the
present invention to a subject under such an administration
condition that transepidermal water loss (TEWL) (g/hm.sup.2)
evaluated in a model animal for skin irritation evaluation is 50 or
less, (ii) administering to a subject the composition providing the
cutaneous TSLP level (pg/mg protein) of 10000 or less evaluated in
a model animal for skin irritation evaluation.
In addition, the composition of the present invention may contain
an additive, if necessary. The additive is selected from, for
example, isotonizing agents, antiseptics/germicides, antioxidants,
resolvents, solubilizers, suspending agents, fillers, pH adjusting
agents, stabilizers, absorption promoters, release rate controlling
agents, coloring agents, plasticizers, crosslinking agents,
adhesives and the like, or a combination of two or more kinds of
them, depending on the compatibility with the main ingredient of
the base, the WT1 peptide and/or the modified WT1 peptide and the
cellular immunity induction promoter, intended administration
regimen and the like. In addition, when the composition of the
present invention is in a tape preparation, the tape preparation
can contain a skin permeability enhancer as an additive.
As used herein, the term "skin permeability enhancer" means any
substance which can improve an efficiency of permeation of a
transdermally administered antigen through the skin, as compared
with the efficiency obtained without the substance. The skin
permeability enhancer is not particularly limited as far as it is
liquid at room temperature (25.degree. C.), that is, has fluidity
at that temperature and has an absorption promoting effect. When
the skin permeability enhancer is a mixture of two or more
substances, the mixture is liquid at room temperature (25.degree.
C.), and has an absorption promoting effect. The skin permeability
enhancer may be an organic liquid and preferably, a hydrophobic
liquid in view of their compatibility with the adhesive layer.
Examples of skin permeability enhancers include higher alcohols
such as oleyl alcohol, octyldodecanol and the like; polyhydric
alcohols such as glycerin, ethylene glycol, polypropylene glycol
and the like; higher fatty acids such as oleic acid, caprylic acid
and the like; fatty acid esters such as isopropyl myristate,
isopropyl palmitate, ethyl oleate and the like; polybasic acid
esters such as diethyl sebacate, diisopropyl adipate and the like;
polyhydric alcohol fatty acid esters such as diglyceryl
triisostearate, monooleic acid sorbitan, dicaprylic acid propylene
glycol, monolauric acid polyethylene glycol, tetraoleic acid
polyoxyethylene sorbit and the like; polyoxyethylene alkyl ethers
such as polyoxyethylene lauryl ether and the like; hydrocarbons
such as squalane, liquid paraffin and the like; plant oils such as
olive oil, castor oil and the like; silicone oils; pyrrolidones
such as N-methylpyrrolidone, N-dodecylpyrrolidone and the like;
sulfoxides such as decylmethyl sulfoxide and the like, and these
can be used alone, or can be used by mixing two or more kinds
thereof.
When the rubber or acrylic adhesive is used, a second skin
permeability enhancer can be used. Specific examples of the second
skin permeability enhancer include polyvinylpyrrolidone,
crospovidone, polypropylene glycol, polyvinyl alcohol, carboxyvinyl
polymer, hydroxypropylcellulose and the like, or a mixture thereof,
but not limited to them. In a preferable aspect, the second skin
permeability enhancer of the present invention is
polyvinylpyrrolidone, crospovidone and/or polypropylene glycol.
From the viewpoint of skin permeability enhancement of the WT1
peptide and/or the modified WT1 peptide, a higher alcohol, more
specifically, a higher alcohol having 8 to 18 (preferably 8 to 14)
carbon atoms, a fatty acid ester, more specifically, a fatty acid
ester of fatty acid having 8 to 18 (preferably 12 to 16) carbon
atoms and a monohydric alcohol having 1 to 18 carbon atoms, a
polyhydric alcohol fatty acid ester or the like, particularly, a
fatty acid ester, particularly, isopropyl myristate, isopropyl
palmitate, or diethyl sebacate is preferably used as the skin
permeability enhancer. The amount of such a skin permeability
enhancer is preferably 0.1% by weight to 70% by weight, more
preferably 1% by weight to 65% by weight, more preferably 5% by
weight to 60% by weight based on the total amount of the adhesive
layer. When the amount of the skin permeability enhancer is 0.1% by
weight or more, the high transdermal absorption promoting effect
can be obtained. When the amount is 70% by weight or less, high
transdermal absorbability can be obtained while suppressing
reduction in an adhesive force or a cohesive force of the whole
adhesive layer and, therefore, this is advantageous.
The therapeutically effective amount of the WT1 peptide and/or the
modified WT1 peptide may widely vary depending on severity of the
disease, age and relative health of a subject as well as other
known factors. In general, satisfactory result may be obtained at
one day dose of about 0.1 .mu.g to 1 g/kg body weight. The
pharmacologically acceptable acid or a pharmacologically acceptable
salt thereof is administered simultaneously with an antigen, or
sequentially, and preferred is simultaneous administration. The
effective amount of the pharmacologically acceptable acid or a
pharmacologically acceptable salt thereof can widely vary depending
on a specific acid or salt to be used, the presence or absence of a
cellular immunity induction promoter to be used together and the
like, and satisfactory result is obtained at 0.01 .mu.g to 1 g/kg
body weight. When the cellular immunity induction promoter is used
together, the cellular immunity induction promoter is administered
simultaneously with the WT1 peptide and/or the modified WT1
peptide, or sequentially, and preferred is simultaneous
administration. The effective amount of the cellular immunity
induction promoter can widely vary depending on a specific cellular
immunity induction promoter to be used, the presence or absence of
other cellular immunity induction promoter and the like, and
satisfactory result is obtained at 0.01 .mu.g to 1 g/kg body
weight. The effective amount of a helper peptide can widely vary
depending on a specific other ingredient to be used, the quantity
thereof and the like, and satisfactory result is obtained at 0.01
.mu.g to 1 g/kg body weight. One day dose may be administered in a
single dose or in several divided portions at several times, such
as two times or more, for example, two, three, four or five times.
The composition may be applied continuously for a period of between
1 minute and 7 days per one administration. The administration
interval is appropriately selected from once every day to one year
(e.g. once per 1 day, once per 2 days, once per 3 days, once per 1
week, once per 2 weeks, once per 1 month, once per 3 months, once
per 6 months, once per 1 year) and longer depending on the state of
the patient, severity of the cancer, whether it is for therapeutic
purpose or preventive purpose. Generally, for the purpose of
treating a patient actually having a severe cancer, the WT1 peptide
and/or the modified WT1 peptide are administered at higher
frequency and a higher dose, and for the preventive purpose for a
patient having no cancer, the WT1 peptide and/or the modified WT1
peptide are administered at lower frequency and a lower dose.
In the present invention, physical irritation means any physical
irritation which gives damage to corneum, including scratch and
scraping. For example, operation of tape stripping which removes
corneum with an adhesive tape or the like, operation of giving
damage to the skin with a cutter, and operation using a microneedle
such as perforation in corneum with a fine needle are also included
in the physical irritation.
Transepidermal water loss means the water amount (g) transpired
from 1 m.sup.2 of keratin per one hour. Transepidermal water loss
can be easily measured with a water loss measuring device in a
short time, and is widely used as an index for evaluating the
damage degree of the skin. Also in the present invention,
transepidermal water loss was used as an index of the physical
irritation level.
TSLP (Thymic stromal lymphopoietin) is one of IL-7-like cytokines
which is produced from keratinocyte of the skin, thymus, and a
mucosal epithelial cells, and is known to be involved in the
maturation of dendritic cells, and the differentiation of T cells.
In the present invention, TSLP was used as an index of the chemical
irritation level which is irritation derived from a drug.
The present invention will be explained in more detail and
specifically below by way of Examples. The present invention is not
limited to the scope of Examples.
EXAMPLES
Cream Formulation
A cream formulation having ingredients of the following Table 2 was
produced. Specifically, Db126 peptide (HLA-A*02 type MHC restricted
peptide) or RYF peptide (HLA-A*24 type MHC restricted peptide) or
AYL peptide (HLA-A*24 type MHC restricted peptide), cellular
immunity induction promoters other than a helper peptide, a helper
peptide, a pharmacologically acceptable acid, and optionally, an
additive were weighed at the amounts explicitly described in Table
2, 15 parts by weight of DMSO was further blended, and a base (base
cream) was added thereto to the total of 100 parts by weight, to
obtain a cream formulation. In cream formulations of Examples or
Comparative Examples in which the blending amounts are specifically
described in Table 2, the blending amounts of respective
ingredients were as described in the table.
Each base cream in Table 2 was prepared by blending and kneading
materials according to the ratio described in the following Table
1.
TABLE-US-00002 TABLE 1 Base cream White vaseline 69.0 wt % Sorbitan
monostearate 0.8 wt % Benzyl alcohol 2.7 wt % Cetanol 2.7 wt %
Stearyl alcohol 4.0 wt % Polysorbate 60 4.0 wt % Concentrated
glycerin 2.7 wt % Water 14.1 wt %
White vaseline, sorbitan monostearate, benzyl alcohol, stearyl
alcohol, Polysorbate 60, concentrated glycerin, and dimethyl
sulfoxide (DMSO) were purchased from Wako Pure Chemical Industries,
Ltd. A TFA salt of Db126 antigen peptide, a TFA salt of RYF
peptide, a TFA salt of AYL peptide, an acetic acid salt of Db126
antigen peptide, Peptide-25 (Pep25) and Peptide-25B (Pep25B) were
chemically synthesized, purified by HPLC and used. Cetanol, and
imiquimod (IMQ) were purchased from Tokyo Chemical Industry Co.,
Ltd. Cyclic di-GMP (c-di-GMP) and cyclic di-AMP (c-di-AMP) were
purchased from Biolog Life Science Institute. Pantoea
bacterium-derived lipopolysaccharide manufactured by MACROPHI Inc.,
Pam.sub.3CSK.sub.4 manufactured by InvivoGen, glucopyranosyl lipid
manufactured by InvivoGen (MPLAs), Zymosan manufactured by Nacalai
Tesque, Inc., Poly(I:C) manufactured by InvivoGen, bropirimine
manufactured by TOCRIS bioscience, R848 manufactured by InvivoGen,
sodium hyaluronate manufactured by Kikkoman Biochemifa Company
(microhyaluronic acid FCH), ODN1826 manufactured by InvivoGen,
pidotimod manufactured by Santa Cruz Biotechnology, Inc., bestatin
manufactured by Wako Pure Chemical Industries, Ltd., levamisole
hydrochloride manufactured by MP Biomedicals, suplatast tosylate
manufactured by TOCRIS bioscience, etodolac manufactured by Wako
Pure Chemical Industries, Ltd., and loxoprofen Na manufactured by
Yoshindo Inc. were respectively used.
The following materials were used.
Imiquimod: Tokyo Chemical Industry Co., Ltd., clofibrate:
manufactured by LKT Laboratories, Inc., fenofibrate: manufactured
by Wako Pure Chemical Industries, Ltd., quercetin: manufactured by
Cayman Chemical Company, berberine (berberine chloride n-hydrate):
manufactured by Wako Pure Chemical Industries, Ltd., noscapine:
manufactured by Wako Pure Chemical Industries, Ltd.,
3,3'-diindolylmethane: manufactured by Wako Pure Chemical
Industries, Ltd., xanthone: manufactured by Wako Pure Chemical
Industries, Ltd., parthenolide: manufactured by Wako Pure Chemical
Industries, Ltd., etodolac: manufactured by Wako Pure Chemical
Industries, Ltd., loxoprofen (loxoprofen Na): manufactured by
Yoshindo Inc., indomethacin: manufactured by Wako Pure Chemical
Industries, Ltd., aspirin: manufactured by Sigma-Aldrich,
diclofenac (diclofenac sodium): manufactured by Wako Pure Chemical
Industries, Ltd., ketoprofen: manufactured by Wako Pure Chemical
Industries, Ltd., celecoxib: manufactured by TOCRIS bioscience,
valdecoxib: manufactured by TOCRIS bioscience, docosahexaenoic
acid: manufactured by Cayman Chemical Company,
2',5'-dideoxyadenosine: manufactured by BIOMOL International,
SCH23390: manufactured by Wako Pure Chemical Industries, Ltd.,
ropinirole (ropinirole HCl): manufactured by Ragactives,
rotigotine: manufactured by STARNASCENS, GW627368X: manufactured by
Cayman Chemical Company, sulprostone: manufactured by manufactured
by Cayman Chemical Company, cloprostenol: manufactured by Wako Pure
Chemical Industries, Ltd., BWA868C: manufactured by Cayman Chemical
Company, RO1138452: manufactured by Cayman Chemical Company,
leukotriene B4: manufactured by Cayman Chemical Company,
montelukast (montelukast sodium): manufactured by LG Life Sciences,
zileuton: manufactured by Toronto Research Chemicals, Inc.,
nicotinic acid: manufactured by Wako Pure Chemical Industries,
Ltd., glycyrrhizic acid (dipotassium glycyrrhizinate): manufactured
by Wako Pure Chemical Industries, Ltd., pirfenidone: manufactured
by TOCRIS bioscience, tranilast: manufactured by Wako Pure Chemical
Industries, Ltd., diphenhydramine (diphenhydramine hydrochloride):
manufactured by Wako Pure Chemical Industries, Ltd., famotidine:
manufactured by Wako Pure Chemical Industries, Ltd., immepip
(immepip dihydrobromide): manufactured by TOCRIS bioscience,
proxyfan: manufactured by TOCRIS bioscience, azelastine (azelastine
hydrochloride): manufactured by LKT Laboratories, Inc., cimetidine:
manufactured by Wako Pure Chemical Industries, Ltd.,
4-methylhistamine: manufactured by TOCRIS bioscience, olanzapine:
manufactured by Wako Pure Chemical Industries, Ltd., yohimbine
(yohimbine hydrochloride): manufactured by Wako Pure Chemical
Industries, Ltd., acetylcholine (acetylcholine chloride):
manufactured by Wako Pure Chemical Industries, Ltd., metergoline
(metergoline phenylmethyl ester): manufactured by TOCRIS
bioscience, clozapine: manufactured by Wako Pure Chemical
Industries, Ltd., sumatriptan: manufactured by MYUNG IN PHARM. CO.,
LTD., zolmitriptan: manufactured by Cipla, tolvaptan: manufactured
by Sigma-Aldrich, desmopressin: manufactured by Sigma-Aldrich,
oxybutynin (oxybutynin hydrochloride): manufactured by
Sigma-Aldrich, pilocarpine (pilocarpine hydrochloride):
manufactured by Wako Pure Chemical Industries, Ltd., tamsulosin
(tamsulosin hydrochloride): manufactured by Cipla, midodrine
(midodrine hydrochloride): manufactured by Wako Pure Chemical
Industries, Ltd., propranolol (propranolol hydrochloride):
manufactured by Wako Pure Chemical Industries, Ltd., xylazine:
manufactured by Wako Pure Chemical Industries, Ltd., novokinin:
manufactured by Sigma-Aldrich, baclofen: manufactured by Tokyo
Chemical Industry Co., Ltd., TRAP-6: manufactured by Bachem,
adenosine diphosphate: manufactured by MP Biomedicals,
somatostatin-14: manufactured by Bachem, GW405833: manufactured by
Sigma-Aldrich, SEW2871: manufactured by Cayman Chemical Company,
trimebutine (trimebutine maleate): manufactured by Tokyo Chemical
Industry Co., Ltd., loperamide (loperamide hydrochloride):
manufactured by Wako Pure Chemical Industries, Ltd., melatonin:
manufactured by LKT Laboratories, Inc., biphenylindanone A:
manufactured by Sigma-Aldrich, L-AP4 (L-2-amino-4-phosphonobutyric
acid): manufactured by Wako Pure Chemical Industries, Ltd.
A composite substrate in which a PET film/PET non-woven fabric
laminate product (area 0.7 cm.sup.2) was stuck to a central portion
of an adhesive tape for fixation so that a PET film side was a tape
side was prepared. A cream formulation (4 mg) was applied on a
non-woven fabric portion of this composite substrate, and this was
used as an administration sample of an immunization test.
Mouse Immunization Test 1 (Cream Formulation)
Mouse immunization test was performed with the cream formulations
using a model animal for immunological evaluation. Evaluation of
the immunity induction level was performed by the ELISPOT method.
Specifically, the hair of the back of the mouse was cut. The mouse
was kept until it recovered from the skin damage due to the hair
cutting. After that, a sample was applied to the back of the mouse
for the predetermined time, and then removed. Then, the mouse was
kept for predetermined days and the level of the antigen-specific
cellular immunity was evaluated. After predetermined days from the
application of the sample, the spleen was isolated and a spleen
cell suspension was prepared. Spleen cells (3.times.10.sup.6
cells/well) and the antigen peptide (100 .mu.M) together with the
culturing medium were placed into a well of an ELISPOT plate on
which an anti-mouse IFN-.gamma. antibody had been immobilized. The
plate was cultured for 20 hours under the condition of 37.degree.
C. and 5% CO.sub.2. The number of the spots representing
IFN-.gamma.-producing cells (spot number/3.times.10.sup.6 cells)
was evaluated by the ELISPOT method. In all cases, 4 mg of the
cream formulation was applied once for 24 hours and the spleen was
isolated 6 days after the completion of the application.
In Example 3, the skin was subjected to tape stripping (TS) ten
times using a DUNPLON tape (NITTO DENKO CS SYSTEM CORPORATION, No.
375) before the application of the sample and, in Examples 4 and 5,
the skin was injured with a microcutter (MICRO FEATHER No. 7330G,
manufactured by FEATHER) and a microneedle (needle length 750
.mu.M, Micro Needle Roller System MR75, manufactured by Ostar
Beauty), respectively before the application.
In addition, the cutaneous TSLP level of the mouse after
administration, transepidermal water loss of the mouse before
administration and the skin permeability of Db126 antigen peptide
and imiquimod were also measured in some groups. A mouse used for
evaluating the TSLP level, transepidermal water loss, and skin
permeability was a C57BL/6 mouse.
(Method of Measuring TSLP Level)
When the application of the sample was completed, the skin of the
back of the mouse was isolated, and the skin was ground using a
homogenizer (Physcotron, Microtec Co., Ltd.) in an extraction
solvent (PBS solution containing protease inhibitor (Protease
Inhibitor Cocktail for general use, Sigma-Aldrich) and 10 .mu.M
indomethacin (manufactured by Wako Pure Chemical Industries,
Ltd.)). The ground skin was centrifuged at 4.degree. C. and 9000 g
for 10 minutes, and the supernatant was recovered. The TSLP amount
in the supernatant was measured by ELISA (Mouse TSLP Quantikine
ELISA Kit, R&D Systems). In addition, the total protein amount
in the supernatant was measured by the BCA method (Pierce BCA
Protein Assay Kit, Thermo Fisher Scientific K.K.), and the TSLP
amount was divided by the total protein amount for
standardization.
(Measurement of Transepidermal Water Loss)
Transepidermal water loss was measured using a portable type
switching chamber system water loss measuring device (manufactured
by Asahibiomed Co., Ltd., VAPO SCAN AS-VT100RS) by contacting a
mouse skin with the device for around 5 to 15 seconds. A value
which was measured 10 minutes after pre-treatment of a mouse skin
was adopted as transepidermal water loss (TEWL) (g/hm.sup.2).
(Mouse Skin Permeability Test)
The skin permeability of Db126 antigen peptide and imiquimod was
determined by using a Franz type diffusion cell. A piece of the
skin was isolated from the back of a mouse, the hair on which had
been cut in advance, was mounted in the Franz-type diffusion cell
(application area 4.91 cm.sup.2) in which phosphate buffer (pH 7.4
isotonic buffer) at 37.degree. C. was circulated. A 0.7 cm.sup.2
preparation was stuck on the mounted skin, and sample in the cell
was collected after 24 hours. The collected sample was subjected to
high performance liquid chromatograph-tandem mass spectrometer, and
the amount of Db126 antigen peptide which had permeated through a
skin after 24 hours (Db126 antigen peptide permeated amount,
.mu.g/cm.sup.2/24 hr) and the amount of imiquimod (imiquimod
permeated amount, .mu.g/cm.sup.2/24 hr) were calculated from a
pre-determined calibration curve.
The results of the immunization test and the results of measurement
of the TSLP level and transepidermal water loss are shown in the
following Table 2 together with the mouse strain used in the
immunization test. The "genetically modified mouse" in Table 2 is a
genetically modified mouse by which cellular immunity induction by
a HLA-A*0201 type MHC restrictive peptide can be evaluated. In
addition, the results of measurement of skin permeability are shown
in Table 3. In addition, for comparison, the results of
immunization using an injectable described later (Comparative
Examples 4 to 8) are described in an end of the table.
TABLE-US-00003 TABLE 2 Results of immunization Composition (ELISPOT
Additive average % Specific Antigen Cellular immunity (Chemical
TSLP Physical TEWL spot Lysis (In Base peptide induction promoter
Acid irritation) (pg/mg protein) irritation (g/h m.sup.2) Mouse
number) vivo CTL assay) Comparative base Db126(25) IMQ(3) PEP(0.3)
None None None 10 C57BL/6 108 - example 1 cream Example 1 base
Db126(25) IMQ(3) PEP(0.3) isostearic None 77 None 10 C57BL/- 6 192
cream acid(8.6) Example 2 base Db126(25) IMQ(3) PEP(0.3) isostearic
SDS(20) 835 None 10 C5- 7BL/6 35 cream acid(6.2) Example 3 base
Db126(25) IMQ(3) PEP(0.3) isostearic None 54 TS 10 times 58 C57BL/6
13 cream acid(8.6) Example 4 base Db126(25) IMQ(3) PEP(0.3)
isostearic None 42 microcutter 66- C57BL/6 8 cream acid(8.6)
Example 5 base Db126(25) IMQ(3) PEP(0.3) isostearic None 53
microneedle 60- C57BL/6 24 cream acid(8.6) Example 6 base Db126(25)
c-di-GMP(cyclic PEP(0.3) isostearic None None 10- C57BL/6 1209 63
cream dinucleotide) acid(8.8) (1) Comparative base RYF(TFA IMQ(4)
PEPB None None None 10 BALB/c 56 example 2 cream salt)(4) (0.3)
Example 7 base RYF(TFA IMQ(4) PEPB isostearic None 77 None 10
BALB/c 81 cream salt)(4) (0.3) acid(8.5) Comparative base AYL(TFA
IMQ(4) PEPB None None None 10 BALB/c 375 example 3 cream salt)(4)
(0.3) Example 8 base AYL(TFA IMQ(4) PEPB isostearic None 78 None 10
BALB/c 471 cream salt)(4) (0.3) acid(8.5) Example 9 base Db126(25)
c-di-GMP(cyclic PEP(0.3) isostearic None None 10- genetically 1500
80 cream dinucleotide) acid(8.8) modified (1) Example 10 base
Db126(25) c-di-AMP(cyclic PEP(0.3) isostearic None None 1- 0
genetically 1498 cream dinucleotide) acid(8.8) modified (1) Example
11 base Db126 Pam3CSK4(TLR1/2 PEP(0.3) isostearic None None 10 ge-
netically 141 cream (25) ligand) acid(8.8) modified (1) Example 12
base Db126 Zymosan(ligand PEP(0.3) isostearic None None 10 gen-
etically 152 cream (25) for TLR2 and acid(8.8) modified Dectin1)
(1) Example 13 base Db126 Poly(I:C) (TLR3 PEP(0.3) isostearic None
None 10 genetically 265 cream (25) ligand) acid(8.8) modified (1)
Example 14 base Db126 lipopolysaccharide PEP(0.3) isostearic None
None 10- genetically 173 84 cream (25) derived from acid(8.8)
modified Pantoea bacterium(TLR4 ligand) (1) Example 15 base Db126
glucopyranosyl PEP(0.3) isostearic None None 10 gen- etically 207
cream (25) lipid(TLR4 acid(8.8) modified ligand) (1) Example 16
base Db126 sodium PEP(0.3) isostearic None None 10 genetically- 114
cream (25) hyaluronate(TLR acid(8.6) modified 4 ligand) (3) Example
17 base Db126 IMQ(3) PEP(0.3) isostearic None None 10 genetically-
342 cream (25) acid(8.6) modified Example 18 base Db126
bropirimine(TLR PEP(0.3) isostearic None None 10 ge- netically 267
cream (25) 7 and/or TLR8 acid(8.8) modified ligand) (1) Example 19
base Db126 R848(TLR7 PEP(0.3) isostearic None None 10 genetica- lly
348 cream (25) and/or TLR8 acid(8.8) modified ligand) (1) Example
20 base Db126 ODN1826(TLR9 PEP(0.3) isostearic None None 10 genet-
ically 405 cream (25) ligand) acid(8.8) modified (1) Example 21
base Db126 pidotimod(immunomodulatory PEP(0.3) isostearic None -
None 10 genetically 67 cream (25) small molecule acid(8.8) modified
drug) (1) Example 22 base Db126 Bestatin(immunomodulatory PEP(0.3)
isostearic None - None 10 genetically 96 cream (25) small molecule
acid(8.8) modified drug) (1) Example 23 base Db126 levamisole
PEP(0.3) isostearic None None 10 genetic- ally 266 cream (25)
hydrochloride(immunomodulatory acid(8.8) modified small molecule
drug) (1) Example 24 base Db126 suplatast PEP(0.3) isostearic None
None 10 genetica- lly 159 cream (25) tosylate(Th2 acid(8.6)
modified cytokine inhibitor)(3) Example 25 base Db126 etodolac(COX
PEP(0.3) isostearic None None 10 genet- ically 325 cream (25)
inhibitor) (3) acid(8.6) modified Example 26 base Db126 loxoprofen
PEP(0.3) isostearic None None 10 genetic- ally 352 cream (25)
Na(COX acid(8.6) modified inhibitor) (3) Example 27 base Db126
clofibrate(PPAR PEP(0.3) isostearic None None 12 ge- netically 315
cream (25) agonist) acid(8.6) modified (3) Example 28 base Db126
fenofibrate(PPA PEP(0.3) isostearic None None 12 ge- netically 237
cream (25) R agonist) (3) acid(8.6) modified Example 29 base Db126
quercetin(TSLP PEP(0.3) isostearic None None 12 gen- etically 156
cream (25) production acid(8.6) modified inhibitor) (3) Example 30
base Db126 berberine(TSLP PEP(0.3) isostearic None None 12 gen-
etically 135 cream (25) production acid(8.6) modified inhibitor)
(3) Example 31 base Db126 noscapine(TSLP PEP(0.3) isostearic None
None 12 gen- etically 355 cream (25) production acid(8.6) modified
inhibitor) (3) Example 32 base Db126 3,3'-diindolylmethane PEP(0.3)
isostearic None None- 12 genetically 324 cream (25) (TSLP acid(8.6)
modified production inhibitor) (3) Example 33 base Db126
xanthone(TSLP PEP(0.3) isostearic None None 12 gene- tically 356
cream (25) production acid(8.6) modified inhibitor) (3) Example 34
base Db126 parthenolide(TSLP PEP(0.3) isostearic None None 12 -
genetically 305 cream (25) production acid(8.6) modified inhibitor)
(3) Example 35 base Db126 indomethacin(COX PEP(0.3) isostearic None
None 12 g- enetically 135 cream (25) inhibitor) (3) acid(8.6)
modified Example 36 base Db126 aspirin(COX PEP(0.3) isostearic None
None 12 geneti- cally 132 cream (25) inhibitor) (3) acid(8.6)
modified Example 37 base Db126 diclofenac(COX PEP(0.3) isostearic
None None 12 gen- etically 245 cream (25) inhibitor) (3) acid(8.6)
modified Example 38 base Db126 ketoprofen(COX PEP(0.3) isostearic
None None 12 gen- etically 165 cream (25) inhibitor) (3) acid(8.6)
modified Example 39 base Db126 celecoxib(COX PEP(0.3) isostearic
None None 12 gene- tically 303 cream (25) inhibitor) (3) acid(8.6)
modified Example 40 base Db126 valdecoxib(COX PEP(0.3) isostearic
None None 12 gen- etically 265 cream (25) inhibitor) (3) acid(8.6)
modified Example 41 base Db126 docosahexaenoic PEP(0.3) isostearic
None None 12 ge- netically 124 cream (25) acid(omega-3 acid(8.6)
modified fatty acid) (3) Example 42 base Db126
2',5'-dideoxyadenosine PEP(0.3) isostearic None Non- e 12
genetically 134 cream (25) (adenylate acid(8.6) modified cyclase
inhibitor) (3) Example 43 base Db126 SCH23390(dopamine PEP(0.3)
isostearic None None ge- netically 110 cream (25) receptor
acid(8.6) modified antagonist) (3) Example 44 base Db126
ropinirole(dopamine PEP(0.3) isostearic None None - genetically 103
cream (25) receptor acid(8.6) modified agonist)(3) Example 45 base
Db126 rotigotine(dopamine PEP(0.3) isostearic None None -
genetically cream (25) receptor acid(8.6) modified agonist) (3)
Example 46 base Db126 GW627368X(prostaglandin PEP(0.3) isostearic
None No- ne 12 genetically 205 cream (25) receptor acid(8.6)
modified antagonist) (3) Example 47 base Db126 sulprostone PEP(0.3)
isostearic None None 12 geneti- cally 231 cream (25) (prostaglandin
acid(8.6) modified receptor agonist) (3) Example 48 base Db126
cloprostenol(prostaglandin PEP(0.3) isostearic None - None 12
genetically 203 cream (25) receptor acid(8.6) modified agonist)(3)
Example 49 base Db126 BWA868C PEP(0.3) isostearic None None
genetically cream (25) (prostaglandin acid(8.6) modified receptor
antagonist) (3) Example 50 base Db126 RO1138452(prost PEP(0.3)
isostearic None None 12 ge- netically 123 cream (25) aglandin
acid(8.6) modified receptor antagonist) (3) Example 51 base Db126
leukotriene PEP(0.3) isostearic None None 12 geneti- cally 132
cream (25) B4(leukotriene acid(8.6) modified receptor agonist) (3)
Example 52 base Db126 montelukast(leukotriene PEP(0.3) isostearic
None No- ne 12 genetically 132 cream (25) receptor acid(8.6)
modified antagonist) (3) Example 53 base Db126 zileuton(leukotriene
PEP(0.3) isostearic None None - 12 genetically 132 cream (25)
receptor acid(8.6) modified antagonist) (3) Example 54 base Db126
nicotinic PEP(0.3) isostearic None None 12 genetica- lly 102 cream
(25) acid(niacin) acid(8.6) modified (adenylate cyclase inhibitor)
(3) Example 55 base Db126 dipotassium PEP(0.3) isostearic None None
12 geneti- cally 107 cream (25) glycyrrhizinate acid(8.6) modified
(phospholipase A2 inhibitor) (3) Example 56 base Db126
pirfenidone(TGF- PEP(0.3) isostearic None None 12 g- enetically 197
cream (25) beta acid(8.6) modified production inhibitor) (3)
Example 57 base Db126 tranilast(TGF-beta PEP(0.3) isostearic None
None 12- genetically 218
cream (25) production acid(8.6) modified inhibitor) (3) Example 58
base Db126 diphenhydramine PEP(0.3) isostearic None None 12 ge-
netically 54 cream (25) (histamine acid(8.6) modified receptor
antagonist) (3) Example 59 base Db126 azelastine(histamine PEP(0.3)
isostearic None None - 12 genetically 286 cream (25) receptor
acid(8.6) modified antagonist) (3) Example 60 base Db126
cimetidine(histamine PEP(0.3) isostearic None None - 12 genetically
187 cream (25) receptor acid(8.6) modified antagonist) (3) Example
61 base Db126 famotidine(histamine PEP(0.3) isostearic None None -
12 genetically 187 cream (25) receptor acid(8.6) modified
antagonist) (3) Example 62 base Db126 proxyfan(histamine PEP(0.3)
isostearic None None g- enetically cream (25) receptor acid(8.6)
modified agonist) (3) Example 63 base Db126 4-methylhistamine
PEP(0.3) isostearic None None 12 - genetically 189 cream (25)
(histamine acid(8.6) modified receptor agonist) (3) Example 64 base
Db126 olanzapine(serotonin PEP(0.3) isostearic None None - 12
genetically 225 cream (25) receptor acid(8.6) modified antagonist)
(3) Example 65 base Db126 yohimbine(serotonin PEP(0.3) isostearic
None None 1- 2 genetically 235 cream (25) receptor acid(8.6)
modified antagonist) (3) Example 66 base Db126
acetylcholine(muscarine PEP(0.3) isostearic None No- ne 12
genetically 102 cream (25) receptor acid(8.6) modified
antagonist)(3) Example 67 base Db126 metergoline(serotonin PEP(0.3)
isostearic None None- genetically cream (25) receptor acid(8.6)
modified antagonist)(3) Example 68 base Db126 clozapine(serotonin
PEP(0.3) isostearic None None 1- 2 genetically 165 cream (25)
receptor acid(8.6) modified antagonist)(3) Example 69 base Db126
sumatriptan(serotonin PEP(0.3) isostearic None None- 12 genetically
523 cream (25) receptor acid(8.6) modified agonist)(3) Example 70
base Db126 zolmitriptan(serotonin PEP(0.3) isostearic None Non- e
12 genetically 154 cream (25) receptor acid(8.6) modified agonist)
(3) Example 71 base Db126 tolvaptan(vasopressin PEP(0.3) isostearic
None None- genetically 114 cream (25) receptor acid(8.6) modified
antagonist) (3) Example 72 base Db126 desmopressin(vasopressin
PEP(0.3) isostearic None N- one 12 genetically 165 cream (25)
receptor acid(8.6) modified agonist) (3) Example 73 base Db126
oxybutynin(muscarine PEP(0.3) isostearic None None - 12 genetically
215 cream (25) receptor acid(8.6) modified antagonist) (3) Example
74 base Db126 pilocarpine(muscarine PEP(0.3) isostearic None None-
12 genetically 134 cream (25) receptor acid(8.6) modified agonist)
(3) Example 75 base Db126 tamsulosin(adrenalin PEP(0.3) isostearic
None None - 12 genetically 203 cream (25) receptor acid(8.6)
modified antagonist) (3) Example 76 base Db126
propranolol(adrenalin PEP(0.3) isostearic None None- genetically
cream (25) receptor acid(8.6) modified antagonist) (3) Example 77
base Db126 xylazine(adrenalin PEP(0.3) isostearic None None 12-
genetically 87 cream (25) receptor acid(8.6) modified agonist) (3)
Example 78 base Db126 novokinin(angiotensin PEP(0.3) isostearic
None None- genetically 106 cream (25) receptor acid(8.6) modified
agonist) (3) Example 79 base Db126 baclofen(GABA PEP(0.3)
isostearic None None geneti- cally 98 cream (25) receptor acid(8.6)
modified agonist) (3) Example 80 base Db126 TRAP-6(thrombin
PEP(0.3) isostearic None None 12 ge- netically 89 cream (25)
receptor acid(8.6) modified agonist) (3) Example 81 base Db126
adenosine PEP(0.3) isostearic None None geneticall- y 95 cream (25)
diphosphate(ADP acid(8.6) modified receptor agonist) (3) Example 82
base Db126 somatostatin-14 PEP(0.3) isostearic None None 12 ge-
netically 89 cream (25) (somatostatin acid(8.6) modified receptor
mouse agonist) (3) Example 83 base Db126 GW405833 PEP(0.3)
isostearic None None 12 genetical- ly 105 cream (25) (cannabinoid
acid(8.6) modified receptor mouse agonist) (3) Example 84 base
Db126 SEW2871(sphingosine-1 PEP(0.3) isostearic None None- 12
genetically 120 cream (25) phosphate acid(8.6) modified receptor
mouse agonist)(3) Example 85 base Db126 trimebutine(muscarine
PEP(0.3) isostearic None None- 12 genetically 489 cream (25)
receptor acid(8.6) modified antagonist) (3) mouse Example 86 base
Db126 loperamide(opioid PEP(0.3) isostearic None None 12 -
genetically 234 cream (25) receptor acid(8.6) modified agonist) (3)
mouse Example 87 base Db126 melatonin(melatonin PEP(0.3) isostearic
None None 1- 2 genetically 204 cream (25) receptor acid(8.6)
modified agonist) (3) mouse Example 88 base Db126 biphenylindanone
PEP(0.3) isostearic None None 12 g- enetically 165 cream (25)
A(metabotropic acid(8.6) modified glutamate mouse receptor agonist)
(3) Example 89 base Db126 L-AP4(metabotropic PEP(0.3) isostearic
None None 12- genetically 238 cream (25) glutamate acid(8.6)
modified receptor mouse agonist) (3) Example 90 base Db126
loxoprofen PEP(0.3) isostearic None None 12 genetic- ally 803 47
cream (25) Na(COX acid(8.6) modified inhibitor) (3), IMQ(3) Example
91 base Db126 berberine(TSLP PEP(0.3) isostearic None None 12 gen-
etically 557 cream (25) production acid(8.6) modified inhibitor)
(3), IMQ(3) Example 92 base Db126 quercetin(TSLP PEP(0.3)
isostearic None None 12 gen- etically 769 46 cream (25) production
acid(8.6) modified inhibitor) (3), mouse IMQ(3) Comparative Saline
Db126 Montanide ISA51VG None None None genetically 33 example 4
(0.033) (50) modified Comparative Saline Db126 Montanide ISA51VG
None None None genetically 28 example 5 (0.1) (50) modified
Comparative Saline Db126 Montanide ISA51VG None None None
genetically 335 example 6 (0.33) (50) modified Comparative Saline
Db126(1) Montanide ISA51VG None None None genetically 347 example 7
(50) modified Comparative Saline Db126 Montanide ISA51VG None None
None genetically 461 32 example 8 (3.3) (50) modified IMQ:
Imiquimod (TLR7 and/or TLR8 ligand) c-di-GMP: Cyclic di-GMP (cyclic
dinucleotide) c-di-AMP: Cyclic di-AMP (cyclic dinucleotide) PEP:
Peptide-25 (SEQ ID No.: 7) (helper peptide) PEPB: Peptide-25B (SEQ
ID No.: 11) (helper peptide) poly(I:C): Polyinosinic-polycytidylic
acid (TLR3 ligand) R848: Resiquimod (TLR7 and/or TLR8 ligand) SDS:
Sodium dodecylsulfate TS: Tape stripping Db126 peptide is in a form
of an acetic acid salt. RYF peptide and AYL peptide are in a form
of TFA salt. A numerical value in parenthesis is blending ratio
(part(s) by weight) of each ingredient (the same in following
Tables).
TABLE-US-00004 TABLE 3 Permeated Db126 Permeated Skin antigen
peptide Imiquimod No. treatment (.mu.g/cm.sup.2/24 hr)
(.mu.g/cm.sup.2/24 hr) Example 3 TS 10 times 282 10 Example 4
Microcutter 271 11 Example 5 Microneedle 179 6 Example 1 None 35
3
Tape Preparation
Adhesives for tape preparation were prepared.
(Polymerization of Acrylic Adhesive A)
Under an inert gas atmosphere, 75 parts of 2-ethylhexyl acrylate,
22 parts of N-vinyl-2-pyrrolidone, 3 parts of acrylic acid and 0.2
part of azobisisobutyronitrile were solution polymerized at
60.degree. C. in ethyl acetate to obtain an acrylic adhesive A
solution.
(Polymerization of Acrylic Adhesive B)
Under an inert gas atmosphere, 70 parts of 2-ethylhexyl acrylate,
25 parts of N-vinyl-2-pyrrolidone, 5 parts of
N-(2-hydroxyethyl)acrylamide and 0.2 part of azobisisobutyronitrile
were solution polymerized at 60.degree. C. in ethyl acetate to
obtain an acrylic adhesive B solution.
(Preparation of PIB Rubber Adhesive)
In toluene were dissolved 24 parts of polyisobutylene (Oppanol
B200, manufactured by BASF), 36 parts of polyisobutylene (Oppanol
B12, manufactured by BASF) and 40 parts of an alicyclic petroleum
resin (Arkon P-100, manufactured by Arakawa Chemical Industries,
Ltd.), to obtain a PIB rubber adhesive solution.
(Preparation of SIS Adhesive A)
In toluene were dissolved 60 parts of a styrene-isoprene-styrene
block copolymer (SIS5002, manufactured by JSR Corporation), and 40
parts of an alicyclic petroleum resin (Arkon P-100, manufactured by
Arakawa Chemical Industries, Ltd.), to obtain a SIS adhesive A
solution.
(Preparation of SIS-PIB Adhesive A)
In toluene were dissolved 30 parts of a styrene-isoprene-styrene
block copolymer (SIS5002, manufactured by JSR Corporation), 30
parts of polyisobutylene (Oppanol B100, manufactured by BASF) and
40 parts of an alicyclic petroleum resin (Arkon P-100, manufactured
by Arakawa Chemical Industries, Ltd.), to obtain a SIS-PIB adhesive
A solution.
Tape preparation shown in the following Table 4 was produced.
Specifically, Db126 antigen peptide, a cellular immunity induction
promoter, a pharmacologically acceptable acid and, optionally, a
skin permeability enhancer, an adhesive solution and an organic
solvent (ethyl acetate, ethanol, toluene etc.) in the amounts
described in Table 4 were blended, kneaded, and spread on a release
liner so that the thickness after drying became about 80 .mu.m, and
the organic solvent was removed by drying, and a support was stuck
to prepare a tape preparation. The adhesive solution was blended so
that the total of respective ingredients and the adhesive after
drying the organic solvent became 100 parts by weight. Polyethylene
terephthalate (PET) film (thickness 25 .mu.m) was used as the
support. Polyethylene terephthalate (PET) sheet (thickness 75
.mu.m) treated with silicone was used as release liner. This tape
preparation was cut to give a piece of 0.7 cm.sup.2, and was
adopted to the immunization test. The release liner was peeled just
before the application of the tape preparation.
Sources from which the Db126 antigen peptide, the cellular immunity
induction promoter and the pharmacologically acceptable acid were
purchased were the same as those in the cream formulation.
Isopropyl myristate (IPM) was purchased from Croda Japan.
WT1.sub.35 (hWT1.sub.35 helper peptide), PADRE (universal helper
peptide) and WT1.sub.332 (WT1.sub.332-347 helper peptide) were
chemically synthesized, and purified by HPLC before use.
Mouse Immunization Test 2 (Tape Preparation)
Using thus obtained tape preparation, mouse immunization test 2 was
performed in the same manner as in the mouse immunization test 1.
The preparation was applied once for 24 hours/week and a spleen was
isolated 6 days after administration. Genetically modified mouse
which can be used to evaluate the cellular immunity inducing
ability of the HLA-A*0201 type MHC restricted peptide was used as
model animal for immunological evaluation.
In Example 69, the skin was injured with a microcutter (MICRO
FEATHER No. 7330G, manufactured by FEATHER) before the application
of the preparation.
In some groups, the cutaneous TSLP level after the application,
transepidermal water loss before the application, and the skin
permeability of Db126 antigen peptide and imiquimod were measured
in the same manner as conducted in the mouse immunization test 1.
The mouse used in evaluation of the TSLP level, transepidermal
water loss, and skin permeability was a C57BL/6 mouse.
The results of the immunization test, and the results of
measurement of the TSLP level and transepidermal water loss are
shown in the following Table 4. In addition, the results of
measurement of skin permeability are shown in Table 5.
TABLE-US-00005 TABLE 4 Results of immunization Composition (ELISPOT
Additive average % Specific Antigen Cellular immunity Skin
permeability (Chemical TSLP Physical TEWL spot Lysis (In Base
peptide induction promoter enhancer Acid irritation) (pg/mg
protein) irritation (g/h m.sup.2) number) vivo CTL assay)
Comparative PIB Db126(10) None None IPM(36) None None None 12 3
example 9 Example 93 PIB Db126(10) None None IPM(27) MA(9) None
None 12 10 Comparative PIB Db126(10) IMQ(1) None IPM(35.6) None
None None 12 25 example 10 Comparative PIB Db126(10) None PEP(1)
IPM(35.6) None None None 12 11 example 11 Comparative PIB Db126(10)
IMQ(1) PEP(1) IPM(35.2) None None None 10 40 example 12 Example 94
PIB Db126(10) None PEP(1) IPM(35.6) MA(8.6) None None 12 18 Example
95 PIB Db126(10) IMQ(1) PEP(1) IPM(25.8) MA(8.6) None 48 None 10 4-
06 Example 96 PIB Db126(10) IMQ(1) PEP(1) IPM(25.2) MA(8.4) BL-4.2
180 None 1- 0 70 (4) Example 97 PIB Db126(10) IMQ(3) PEP(1)
IPM(25.8) MA(8.6) None None 10 584- 35 Example 98 PIB Db126(10)
IMQ(3) WT1.sub.35(1) IPM(25.8) MA(8.6) None None- 10 1113 55
Example 99 PIB Db126(10) IMQ(3) WT1.sub.35 IPM(25.8) MA(8.6) None
T/S10 5- 8 56 (1) Example 100 PIB Db126(10) IMQ(3) WT1.sub.35(1)
IPM(25.8) MA(8.6) None mic- rocutter 66 29 Comparative Acryl A
Db126(10) None None IPM(26.4) None None None 10 2 example 13
Example 101 Acryl A Db126(10) None None IPM(26.4) MA(8.8) None None
10 11 Example 102 Acryl A Db126(10) IMQ(1) None IPM(26.4) MA(8.8)
None None 10 31 Example 103 Acryl A Db126(10) IMQ(1) PEP(1)
IPM(26.4) MA(8.8) None None 10 49 Example 104 Acryl B Db126(10)
IMQ(1) PEP(1) IPM(26.4) MA(8.8) None None 10 75 Comparative SIS
Db126(10) IMQ(1) PEP(1) IPM(17.4), None None None 10 30 example 14
Liquid paraffin (17.4) Example 105 SIS Db126(10) IMQ(1) PEP(1)
IPM(13.2), MA(8.4) None None 10 3- 77 Liquid paraffin (13.2)
Comparative SIS-PIB Db126(10) IMQ(1) PEP(1) IPM(17.4), None None
None 10 - 26 example 15 Liquid paraffin (17.4) Example 106 SIS-PIB
Db126(10) IMQ(1) PEP(1) IPM(13.2), MA(8.4) None None - 10 277
Liquid paraffin (13.2) Example 107 PIB Db126(10) IMQ(3) PEP(1)
IPM(33.0) octanoic None None 10 2- 5 acid(1.7) Example 108 PIB
Db126(10) IMQ(3) PEP(1) IPM(33.0) isostearic None None 10- 146
acid(1.7) Example 109 PIB Db126(10) IMQ(3) PEP(1) IPM(25.8)
decanoic None None 10 3- 75 acid(8.6) Example 110 PIB Db126(10)
IMQ(3) PEP(1) IPM(25.8) lauric None None 10 446- acid(8.6) Example
111 PIB Db126(10) IMQ(3) PEP(1) IPM(25.8) palmitic None None 10 4-
0 acid (8.6) Example 112 PIB Db126(10) IMQ(3) PEP(1) IPM(25.8)
isostearic None None 10- 494 acid (8.6) Example 113 PIB Db126(10)
IMQ(3) PEP(1) IPM(25.8) oleic None None 10 495 acid(8.6) Example
114 PIB Db126(10) IMQ(3) PEP(1) IPM(25.8) stearic None None 10 24-
acid (8.6) Example 115 PIB Db126(10) IMQ(3) PEP(1) IPM(31.4) lactic
None None 12 501- acid(3) Example 116 PIB Db126(10) c-di-GMP PEP(1)
IPM(25.8) MA(8.6) None None 45- 0 (0.3) Example 117 PIB Db126(10)
c-di-AMP PEP(1) IPM(25.8) MA(8.6) None None (0.3) Example 118 PIB
Db126(10) lipopolysaccharide PEP(1) IPM(25.8) MA(8.6) None- None 12
103 24 derived from Pantoea bacterium (TLR4 ligand) (3) Example 119
PIB Db126(10) glucopyranosyl PEP(1) IPM(25.8) MA(8.6) None No- ne
lipid(TLR4 ligand) (3) Example 120 PIB Db126(10) sodium hyaluronate
PEP(1) IPM(25.8) MA(8.6) None None (TLR4 ligand) (3) Exdiple 121
PIB Db126(10) ODN1826(TLR9 PEP(1) IPM(25.8) MA(8.6) None None-
ligand) (3) Exanple 122 PIB Db126(10) levamisole PEP(1) IPM(25.8)
MA(8.6) None None 1- 2 45 hydrochloride(immunomodulatory small
molecule drug) (3) Example 123 PIB Db126(10) etodolac(COX PEP(1)
IPM(25.8) MA(8.6) None None- inhibitor) (3) Example 124 PIB
Db126(10) pidotimod(immunomodulatory PEP(1) IPM(25.8) MA(8- .6)
None None small molecule drug) (3) Example 125 PIB Db126(10)
Bestatin(immunomodulatory PEP(1) IPM(25.8) MA(8.- 6) None None
small molecule drug) (3) Example 126 PIB Db126(10) None PEP(1)
IPM(25.8) MA(8.6) None None 10 41 Comparative PIB Db126(10)
loxoprofen Na(COX PEP(1) IPM(25.8) None None None 10 185 40 example
16 inhibitor) (3) Example 127 PIB Db126(10) loxoprofen Na(COX
PEP(1) IPM(25.8) MA(8.6) None None 10 610 inhibitor) (3) Example
128 PIB Db126(10) loxoprofen Na(COX PEP(1) IPM(25.8) MA(8.6) None
None 10 736 40 inhibitor) (1.5), IMQ(1.5) Example 129 PIB Db126(10)
quercetin(TSLP PEP(1) IPM(25.8) MA(8.6) None No- ne 12 560
production inhibitor) (3) Example 130 PIB Db126(10) quercetin(TSLP
PEP(1) IPM(25.8) MA(8.6) None No- ne 12 621 35 production
inhibitor) (1.5), IMQ(1.5) Example 131 PIB Db126
GW627368X(prostaglandin PEP(1) IPM(25.8) MA(8.6) Non- e None 12 574
(10) receptor antagonist) (3) Example 132 PIB Db126 sulprostone
PEP(1) IPM(25.8) MA(8.6) None None 12 5- 30 (10) (prostaglandin
receptor agonist) (3) Example 133 PIB Db126 clofibrate(PPAR PEP(1)
IPM(25.8) MA(8.6) None None - 12 610 (10) agonist) (3) Example 134
PIB Db126 tranilast(TGF-beta PEP(1) IPM(25.8) MA(8.6) None No- ne
12 123 (10) production inhibitor) (3) Example 135 PIB Db126
Immepip(histamine PEP(1) IPM(25.8) MA(8.6) None Non- e 12 155 (10)
receptor agonist) (3) Example 136 PIB Db126 azelastine(histamine
PEP(1) IPM(25.8) MA(8.6) None - None 12 770 (10) receptor
antagonist) (3) Example 137 PIB Db126 sumatriptan(serotonin PEP(1)
IPM(25.8) MA(8.6) None - None 12 790 (10) receptor agonist) (3)
Example 138 PIB Db126 yohimbine(serotonin PEP(1) IPM(25.8) MA(8.6)
None N- one 12 173 (10) receptor antagonist) (3) Example 139 PIB
Db126 oxybutynin(muscarine PEP(1) IPM(25.8) MA(8.6) None - None 12
185 (10) receptor antagonist) (3) Example 140 PIB Db126
tamsulosin(adrenal PEP(1) IPM(25.8) MA(8.6) None No- ne 12 171 (10)
in receptor antagonist) (3) Example 141 PIB Db126 loperamide
(opioid PEP(1) IPM(25.8) MA(8.6) None None 12 480 (10) receptor
agonist) (3) Example 142 PIB Db126 melatonin (melatonin PEP(1)
IPM(25.8) MA (8.6) None None 12 535 (10) receptor agonist) (3)
Example 143 PIB Db126 L-AP4(metabotropic PEP(1) IPM(25.8) MA(8.6)
None No- ne 12 111 (10) glutamate receptor agonist) (3) Example 144
PIB Db126(10) IMQ(3) PADRE IPM(25.8) MA(8.6) None None 12 613- (1)
Example 145 PIB Db126(10) IMQ(3) WT1.sub.332 IPM(25.8) MA(8.6) None
None (1) Example PIB Db126(10) IMQ(3) PEP(1) IPP(25.8) MA(8.6) None
None 12 560 146 Example PIB Db126(10) IMQ(3) PEP(1) None MA(8.6)
None None 12 54 147 Comparative Saline Db126 Montanide ISA51VG None
None None None 33 example 4 (0.033) (50) Comparative Saline Db126
Montanide ISA51VG None None None None 28 example 5 (0.1) (50)
Comparative Saline Db126 Montanide ISA51VG None None None None 335
example 6 (0.33) (50) Comparative Saline Db126(1) Montanide ISA51VG
None None None None 347 example 7 (50) Comparative Saline Db126
Montanide ISA51VG None None None None 461 32 example 8 (3.3) (50)
Acryl A: Acrylic adhesive A Acryl B: Acrylic adhesive B PIB: PIB
rubber adhesive SIS: SIS adhesive A SIS-PIB: SIS-PIB adhesive A
WT1.sub.35: hWT1.sub.35 helper peptide (SEQ ID No.: 8) (helper
peptide) PADRE: Universal helper peptide (SEQ ID No.: 9) (helper
peptide) WT1.sub.332: WT1.sub.332-347 helper peptide (SEQ ID No.:
10) (helper peptide) IPM: Isopropyl myristate, manufactured by
Croda Japan IPP: Isopropyl palmitate, manufactured by Wako Pure
Chemical Industries, Ltd. MA: Myristic acid BL-4.2: Polyoxyethylene
(4,2) lauryl ether, manufactured by Nikko Chemicals Co., Ltd. Db126
peptide is in a form of an acetic acid salt, in all cases. A
numerical value in parenthesis is blending ratio (part(s) by
weight) of each ingredient.
TABLE-US-00006 TABLE 5 Permeated Db126 Permeated antigen peptide
Imiquimod No. Characteristic (.mu.g/cm.sup.2/24 hr)
(.mu.g/cm.sup.2/24 hr) Example 95 Surfactant-free 2.8 4.5 tape
Example 96 BL-4.2 containing 13.4 5.8 tape
Liquid Formulation for External Use
A liquid formulation for external use having a composition of the
following Table 6 was produced. Db126 antigen peptide, a cellular
immunity induction promoter and a pharmacologically acceptable acid
at the blending amounts described in Table 6, and 15 parts by
weight of DMSO were blended, a base was added thereto to the total
of 100 parts by weight, and the materials were kneaded to obtain a
liquid formulation for external use. Propylene glycol (PG) and
oleyl alcohol (OA) were mixed so that the weight ratio became the
ratio of 98:2 or 90:10 to give the base. A composite substrate in
which a cellulose non-woven fabric portion (area 0.8 cm.sup.2) was
stuck to a central portion of an adhesive tape for fixation was
prepared. The non-woven fabric portion of this composite substrate
was impregnated with 67 .mu.L of the prepared liquid formulation
for external use, and this was used in the immunization test.
Sources from which the Db126 antigen peptide, the cellular immunity
induction promoter and the pharmacologically acceptable acid were
purchased were the same as those in the cream formulation. Lactic
acid was purchased from Wako Pure Chemical Industries, Ltd.
Mouse Immunization Test 3 (Liquid Formulation for External Use)
Using a liquid formulation for external use produced as described
above, mouse immunization test was performed in the same manner as
that in the mouse immunization test 1. The dose was 67 .mu.L as
described above. The liquid formulation was applied once for 24
hours/week and a spleen was isolated 6 days after completion of the
application. The mouse used was a C57BL/6.
In some groups, the cutaneous TSLP level of amuse after the
application, and transepidermal water loss of a mouse before the
application were measured using a C57BL/6 mouse in the same manner
as conducted in the mouse immunization test 1.
The results of the immunization test, the TSLP level and
transepidermal water loss are shown in the following Table 6
together with each mouse used.
TABLE-US-00007 TABLE 6 Result of immuniza- tion (ELISPOT
Composition TSLP average Antigen Cellular immunity Chemical (pg/mg
Physical TEWL spot Base peptide induction promoter Acid irritation
protein) irritation (g/h m.sup.2) Mouse number) Comparative PG/OA
Db126(10) IMQ(10) PEP(0.3) None None 174 None 10 C57BL/6- 503
example 17 [98/2] Exanple 148 PG/OA Db126(10) IMQ(10) PEP(0.3)
lactic None 185 None 10 C57BL- /6 595 [98/2] acid(0.5) Example 149
PG/OA Db126(10) IMQ(10) PEP(0.3) salicylic None None 10 C57BL- /6
576 [98/2] acid(0.5) Example 150 PG/OA Db126(10) IMQ(10) PEP(0.3)
DL-malic None None 10 C57BL/- 6 520 [98/2] acid(0.5) Example 151
PG/OA Db126(10) IMQ(10) PEP(0.3) methanesulfonic None None 10-
C57BL/6 540 [98/2] acid(0.5) Example 152 PG/OA Db126(10) IMQ(10)
PEP(0.3) maleic None None 10 C57BL/6 - 523 [98/2] acid(0.5) Example
153 PG/OA Db126(10) IMQ(10) PEP(0.3) citric None None 10 C57BL/6 -
552 [98/2] acid(0.5) Comparative PG/OA Db126(10) IMQ(10) PEP(0.3)
None Given 270 None 10 C57BL/- 6 435 example 18 [90/10] (Increase
in OA ratio) Example 154 PG/OA Db126(10) IMQ(10) PEP(0.3) lactic
Given 313 None 10 C57B- L/6 515 [90/10] acid(0.5) (Increase in OA
ratio) PG/OA: A mixture of propylene glycol and oleyl alcohol (both
are manufactured by Wako Pure Chemical Industries, Ltd.). A
numerical value in brackets [ ] represents the amount ratio of PG
and OA. A Db126 peptide is in a form of an acetic acid salt. A
numerical value in parenthesis ( ) is blending ratio (part(s) by
weight) of each ingredient.
Injectable
Intradermal injectable formulations having a composition of the
following Table 7 was produced. Specifically, a saline as a base
was added to a Db126 antigen peptide and Montanide ISA51VG
(manufactured by Freund Corporation) as an adjuvant at the blending
amounts described in Table 7, to the total of 100 parts by weight,
and the materials were kneaded with a homogenizer to prepare an
injectable.
Mouse Immunization Test 4 (Injectable)
Using an injectable produced as described above, mouse immunization
test was performed in the same manner as in the mouse immunization
test 1. 30 .mu.L of the injectable formulation was administered
once to the back of mouse by intradermal injection, and a spleen
was isolated 6 days after the administration. Genetically modified
mouse which can be used to evaluate the cellular immunity inducing
ability of the HLA-A*0201 type MHC restricted peptide was used as
model animal. The results of the immunization test are shown in the
following Table 7.
TABLE-US-00008 TABLE 7 Immunity % result Specific Cellular (ELISPOT
Lysis immunity average (In vivo Antigen induction spot CTL Base
peptide promoter number) assay) Comparative Saline Db126(0.033)
Montanide 33 Example 4 ISA51VG (50) Comparative Saline Db126(0.1)
Montanide 28 Example 5 ISA51VG (50) Comparative Saline Db126(0.33)
Montanide 335 Example 6 ISA51VG (50) Comparative Saline Db126(1)
Montanide 347 Example 7 ISA51VG (50) Comparative Saline Db126(3.3)
Montanide 461 32 Example 8 ISA51VG (50) A Db126 peptide is in a
form of an acetic acid salt in all cases. A numerical value in
parenthesis is blending ratio (part(s) by weight) of each
ingredient.
In Vivo CTL Assay
Seven days after final immunization, the spleen cells (target cell
or control cell) were transplanted according to the following
procedure, the spleen was isolated after 18 hours. The % Specific
Lysis was obtained by performing the FACS measurement.
Procedure 1. Collection of Spleen Cell of Naive Mouse
Naive mouse that is the same kind mouse as that used in the
immunization test was used. Spleen was isolated from the naive
mouse and mashed using a glass slide in a petri dish containing
RPMI1640 medium. The mashed spleen was put into a 50 mL tube and
centrifuged at 10.degree. C. and 1100 rpm for 5 minutes. The
supernatant was discarded. 20 mL of Lysis Buffer was added to the
tube, followed by incubation at room temperature for 5 minutes. 20
mL of the medium was added to the tube and the tube was then
centrifuged. The medium was added to the tube and the resultant was
passed through a cell strainer to give spleen cell suspension.
Procedure 2. Labeling of Spleen Cell with Antigen
The spleen cells prepared in Procedure 1 were centrifuged at
10.degree. C. and 1100 rpm for 5 minutes, the supernatant was
discarded, and HBSS buffer was added to give cell suspension of
2.times.10.sup.7 cells/mL. The cell suspension was dispensed into
two 50 mL tubes, 100 .mu.M of the antigen solution (the antigen was
the same antigen used in the immunization test) was added to one of
the tubes containing the cell solution so that the final
concentration became 10 .mu.M, to obtain a target cell. The cell in
another tube was adopted as control. The cells in both tubes were
incubated at 37.degree. C. for 1 hour, centrifuged, the supernatant
was discarded, and a medium was added.
Procedure 3. Labelling of Spleen Cell with CFSE
The cell labelled with the antigen according to the procedure 2 was
centrifuged, and 0.1% BSA-PBS was added to 1.times.10.sup.7
cells/mL. To the target cell solution was added a 5 mM CFSE
solution to the final concentration of 10 .mu.M, and to the control
cell solution was added a 5 mM CFSE solution to the final
concentration of 1 .mu.M, and the mixture was vortexed, followed by
incubation at 37.degree. C. for 10 minutes. Thereafter,
centrifugation was performed, the supernatant was discarded, and a
medium was added.
Procedure 4. Transplantation of Spleen Cell
The cell labelled with CFSE according to the procedure 3 was
centrifuged, the supernatant was discarded, and cells were adjusted
to 5.times.10.sup.7 cells/mL using a HBSS buffer. Equal amounts of
the target cell solution and the control cell solution were mixed,
and each 200 .mu.L was administered to an immunized mouse via
orbital veins (transplantation cell number: 1.times.10.sup.7
cells/animal).
Procedure 5. Preparation of Spleen Cell of Immunized Mouse and
Measurement of FACS
Eighteen hours after transplantation of the spleen cells, spleen of
the mouse was isolated, and spleen cell suspension was prepared in
the same manner as in Procedure 1. Thereafter, a CFSE-positive cell
was detected by FACS, and the ratio between a CFSE high cell
(target cell) and a CFSE low cell (control cell) was obtained. The
cytotoxic activity was evaluated by the formula shown below. The
obtained value can be used as an index showing the ability of the
antigen-specific killer cells induced by the immunization with the
vaccine composition to specifically attack the cells that present
the antigen in the living body. It was confirmed that the
composition of the present invention can induce strong
antigen-induced cellular immunity. r=(% CFSE low cells)/(% CFSE
high cells) % Specific Lysis=(1-(r_.sub.non
immunized/r_.sub.immunized)).times.100
By comparison between Comparative Example 1 and Example 1,
comparison between Comparative Example 2 and Example 7, comparison
between Comparative Example 3 and Example 8, comparison between
Comparative Example 12 and Example 95, comparison between
Comparative Example 14 and Example 105, comparison between
Comparative Example 15 and Example 106, comparison between
Comparative Example 16 and Example 127, comparison between
Comparative Example 17 and Examples 148-153, and comparison between
Comparative Example 18 and Example 154, it is indicated that
cellular immunity is induced and promoted by addition of an
acid.
In addition, it is indicated that cellular immunity was induced in
various tape preparations with an acid blended therein (see Table 4
and FIGS. 1 and 2). It is indicated that octanoic acid, isostearic
acid, decanoic acid, lauric acid, palmitic acid, isostearic acid,
oleic acid, stearic acid and lactic acid are preferable and that
strong cellular immunity was induced by blending, particularly,
decanoic acid, lauric acid, myristic acid, isostearic acid, oleic
acid and lactic acid. Among them, particularly, lauric acid,
myristic acid, isostearic acid, oleic acid and lactic acid produced
excellent results.
Also in the transdermal immunization using liquid formulation, the
addition of an acid promoted the induction of immunity. Lactic
acid, salicylic acid, malic acid, methanesulfonic acid, maleic
acid, and citric acid were effective. Particularly, lactic acid,
salicylic acid, citric acid, and methanesulfonic acid were
effective.
Further, it is also indicated that cellular immunity stronger than
that by injection was induced (see Tables 4 and 7).
Based on the above results, it was confirmed that a cancer vaccine
composition for transdermal administration for cellular immunity
induction comprising (i) WT1 peptide and/or modified WT1 peptide
and (ii) a pharmacologically acceptable acid or a pharmacologically
acceptable salt thereof as a first cellular immunity induction
promoter was effective for the induction of cellular immunity.
In addition, from Table 2, Table 4 and Table 6, it is also
indicated that cellular immunity is weakened by giving physical
damage to a skin by pre-treatment such as tape stripping or skin
gash, or by giving chemical irritation to a skin by the use of a
surfactant or a specific kind of base material.
Concerning tape preparations in which the kinds of organic fatty
acids were different, clear correspondence was not seen between the
skin permeation amount and the immunity induction level. In
addition, it was confirmed that the skin permeation amount of Db126
and imiquimod was increased by a pre-treatment of a skin.
When compared and checked with the result of an immunity test, it
is indicated that TSLP which is an index of skin irritation was
increased, and the cellular immunity induction level was reduced by
increasing the OA ratio of a PG/OA liquid formulation, or adding a
surfactant to a tape. From this result, it is indicated that it is
important to induce cellular immunity under the mildly irritating
condition.
It is indicated that the TEWL value was increased by pre-treatment
of a skin, and skin barrier was destructed by physical irritation.
This result corresponds to the fact that the skin permeation amount
of Db126 and imiquimod was considerably increased by pre-treatment
of a skin, in the skin permeability test. However, in spite of
increase in the permeation amount, cellular immunity was not
increased, but rather attenuated. From this result, it is indicated
that administration under the mildly irritating condition is more
important for cellular immunity induction rather than increase in
the skin permeation amount of an antigen and an adjuvant.
SEQUENCE LISTINGS
1
1319PRTHomo sapiens 1Arg Met Phe Pro Asn Ala Pro Tyr Leu 1 5
29PRTHomo sapiens 2Cys Met Thr Trp Asn Gln Met Asn Leu 1 5
39PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 3Cys Tyr Thr Trp Asn Gln Met Asn Leu 1 5
49PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 4Arg Tyr Phe Pro Asn Ala Pro Tyr Leu 1 5
59PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 5Ala Tyr Leu Pro Ala Val Pro Ser Leu 1 5
620DNAUnknownDescription of Unknown Bacterial DNA sequence
6tccatgacgt tcctgacgtt 20715PRTMycobacterium tuberculosis 7Phe Gln
Asp Ala Tyr Asn Ala Ala Gly Gly His Asn Ala Val Phe 1 5 10 15
818PRTHomo sapiens 8Trp Ala Pro Val Leu Asp Phe Ala Pro Pro Gly Ala
Ser Ala Tyr Gly 1 5 10 15 Ser Leu 913PRTArtificial
SequenceDescription of Artificial Sequence Synthetic
peptideMOD_RES(1)..(1)D-AlaMOD_RES(3)..(3)CyclohexylalanineMOD_RES(13)..(-
13)D-Ala 9Ala Lys Ala Val Ala Ala Trp Thr Leu Lys Ala Ala Ala 1 5
10 1016PRTHomo sapiens 10Lys Arg Tyr Phe Lys Leu Ser His Leu Gln
Met His Ser Arg Lys His 1 5 10 15 1115PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 11Phe
Gln Asp Ala Tyr Asn Ala Val His Ala Ala His Ala Val Phe 1 5 10 15
125PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 12Ser Lys Lys Lys Lys 1 5 139PRTArtificial
SequenceDescription of Artificial Sequence Synthetic peptide 13Gly
Asp Pro Lys His Pro Lys Ser Phe 1 5
* * * * *